2024-03-29T08:53:03Zhttp://repository.helmholtz-hzi.de/oai/requestoai:repository.helmholtz-hzi.de:10033/3018082019-08-30T11:34:48Zcom_10033_311308col_10033_620561
Hofmeyer, Thomas
0202ea624e946c04ff2f8650fe0e90d9
500
Schmelz, Stefan
91c14d4d519c8fc0f1446b2262626ea5
500
Degiacomi, Matteo T
cfa84a024a3ad25ea53bf63d1a898676
500
Dal Peraro, Matteo
dcb8dddb614d18b8c04d9a3285833fc1
500
Daneschdar, Matin
30be809283a7999b7c7e927d159b4379
500
Scrima, Andrea
c3ad854d78113b599fa24cb5e9e6fbcc
500
van den Heuvel, Joop
3640e46477a8788e4e1d4297adec174d
500
Heinz, Dirk W
31d9fb437708fb56e485ef62c64d3d17
500
Kolmar, Harald
99594f33693a836f3064a4ab58294d07
600
http://orcid.org/0000-0002-8210-1993
Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Petersenstraße 22, 64287 Darmstadt, Germany.
2013-09-18T14:16:21Z
2013-09-18T14:16:21Z
2013-04-26
Arranged sevenfold: structural insights into the C-terminal oligomerization domain of human C4b-binding protein. 2013, 425 (8):1302-17 J. Mol. Biol.
1089-8638
23274142
10.1016/j.jmb.2012.12.017
http://hdl.handle.net/10033/301808
Journal of molecular biology
The complement system as a major part of innate immunity is the first line of defense against invading microorganisms. Orchestrated by more than 60 proteins, its major task is to discriminate between host cells and pathogens and to initiate immune response. Additional recognition of necrotic or apoptotic cells demands a fine-tune regulation of this powerful system. C4b-binding protein (C4BP) is the major inhibitor of the classical complement and lectin pathway. The crystal structure of the human C4BP oligomerization domain in its 7α isoform and molecular simulations provide first structural insights of C4BP oligomerization. The heptameric core structure is stabilized by intermolecular disulfide bonds. In addition, thermal shift assays indicate that layers of electrostatic interactions mainly contribute to the extraordinary thermodynamic stability of the complex. These findings make C4BP a promising scaffold for multivalent ligand display with applications in immunology and biological chemistry.
en
Archived with thanks to Journal of molecular biology
Complement C4b-Binding Protein
Crystallography, X-Ray
Humans
Models, Molecular
Molecular Dynamics Simulation
Protein Conformation
Protein Multimerization
Static Electricity
Thermodynamics
Arranged sevenfold: structural insights into the C-terminal oligomerization domain of human C4b-binding protein.
Article
2018-06-13T19:27:11Z
The complement system as a major part of innate immunity is the first line of defense against invading microorganisms. Orchestrated by more than 60 proteins, its major task is to discriminate between host cells and pathogens and to initiate immune response. Additional recognition of necrotic or apoptotic cells demands a fine-tune regulation of this powerful system. C4b-binding protein (C4BP) is the major inhibitor of the classical complement and lectin pathway. The crystal structure of the human C4BP oligomerization domain in its 7α isoform and molecular simulations provide first structural insights of C4BP oligomerization. The heptameric core structure is stabilized by intermolecular disulfide bonds. In addition, thermal shift assays indicate that layers of electrostatic interactions mainly contribute to the extraordinary thermodynamic stability of the complex. These findings make C4BP a promising scaffold for multivalent ligand display with applications in immunology and biological chemistry.
CC-LICENSE
license_url
license_url
text/plain
49
https://hzi.openrepository.com/bitstream/10033/301808/3/license_url
c0b3e2a72fe5e4b72fa5798bb22cd3a3
MD5
3
false
license_text
license_text
application/octet-stream
0
https://hzi.openrepository.com/bitstream/10033/301808/4/license_text
d41d8cd98f00b204e9800998ecf8427e
MD5
4
false
license_rdf
license_rdf
application/octet-stream
24332
https://hzi.openrepository.com/bitstream/10033/301808/5/license_rdf
306824103cdab5a2460a7737d9c97e69
MD5
5
false
LICENSE
license.txt
license.txt
text/plain
1685
https://hzi.openrepository.com/bitstream/10033/301808/6/license.txt
cb598eeb10bfed09d26fd8d285172ad4
MD5
6
false
ORIGINAL
Hofmeyer et al_final.pdf
Hofmeyer et al_final.pdf
original manuscript with figures
application/pdf
1843098
https://hzi.openrepository.com/bitstream/10033/301808/1/Hofmeyer%20et%20al_final.pdf
44ced98497d2cb17d2c27b5dabb33f6e
MD5
1
true
Hofmeyer et al 2013 supplemental.pdf
Hofmeyer et al 2013 supplemental.pdf
supplemental figures
application/pdf
1674807
https://hzi.openrepository.com/bitstream/10033/301808/2/Hofmeyer%20et%20al%202013%20supplemental.pdf
6ac54e6a81c6228536616de500cd6af1
MD5
2
false
TEXT
Hofmeyer et al 2013 supplemental.pdf.txt
Hofmeyer et al 2013 supplemental.pdf.txt
Extracted Text
text/plain
14361
https://hzi.openrepository.com/bitstream/10033/301808/11/Hofmeyer%20et%20al%202013%20supplemental.pdf.txt
d5a5e6d5387a77dcc8e4ea3bab763469
MD5
11
false
Hofmeyer et al_final.pdf.txt
Hofmeyer et al_final.pdf.txt
Extracted Text
text/plain
57807
https://hzi.openrepository.com/bitstream/10033/301808/12/Hofmeyer%20et%20al_final.pdf.txt
48150b9bdad6b3d0c2bd0a8980bd93a8
MD5
12
false
THUMBNAIL
Hofmeyer et al_final.pdf.jpg
Hofmeyer et al_final.pdf.jpg
Generated Thumbnail
image/jpeg
33515
https://hzi.openrepository.com/bitstream/10033/301808/14/Hofmeyer%20et%20al_final.pdf.jpg
dec4097dad9535ce4cd9ce6e1f57be51
MD5
14
false
Hofmeyer et al 2013 supplemental.pdf.jpg
Hofmeyer et al 2013 supplemental.pdf.jpg
Generated Thumbnail
image/jpeg
13108
https://hzi.openrepository.com/bitstream/10033/301808/15/Hofmeyer%20et%20al%202013%20supplemental.pdf.jpg
acea59f8d83e060566288d38df2cc637
MD5
15
false
elsevier-thumbnail.png
application/octet-stream
114958
https://hzi.openrepository.com/bitstream/10033/301808/16/elsevier-thumbnail.png
a2388eb93f6d523dcef027aaabdf5d50
MD5
16
false
10033/301808
oai:hzi.openrepository.com:10033/301808
2019-08-30 11:34:48.448
Helmholtz Zentrum für Infektionsforschung Repository
hzi@openrepository.com
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
oai:repository.helmholtz-hzi.de:10033/6207832018-06-13T00:40:48Zcom_10033_311308col_10033_620561
He, Feng Q
76fe8ad1-77a4-4d65-84a7-001d9f16c146
-1
Sauermann, Ulrike
2872b74e-02c5-4e7b-a5e0-e82c655511f8
-1
Beer, Christiane
a8d62547-a0c0-4ad8-9345-79490e5d9bb7
-1
Winkelmann, Silke
c450a866-323f-4b3f-827e-b022d5f89892
-1
Yu, Zheng
4c5f47af-a357-4087-abf7-f3f9ca1ea90e
-1
Sopper, Sieghart
d8f1b27c-3781-4c25-a00b-27a9f37af8a2
-1
Zeng, An-Ping
0000-0001-9768-7096
600
Wirth, Manfred
8363031d-909e-4f38-ba0f-803a61db32c6
-1
2017-01-27T11:36:19Z
2017-01-27T11:36:19Z
2014-08-27
2015-09-04T08:27:08Z
Virology Journal. 2014 Aug 27;11(1):152
http://dx.doi.org/10.1186/1743-422X-11-152
http://hdl.handle.net/10033/620783
Abstract Background The deciphering of cellular networks to determine susceptibility to infection by HIV or the related simian immunodeficiency virus (SIV) is a major challenge in infection biology. Results Here, we have compared gene expression profiles of a human CD4+ T cell line at 24 h after infection with a cell line of the same origin permanently releasing SIVmac. A new knowledge-based-network approach (Inter-Chain-Finder, ICF) has been used to identify sub-networks associated with cell survival of a chronically SIV-infected T cell line. Notably, the method can identify not only differentially expressed key hub genes but also non-differentially expressed, critical, ‘hidden’ regulators. Six out of the 13 predicted major hidden key regulators were among the landscape of proteins known to interact with HIV. Several sub-networks were dysregulated upon chronic infection with SIV. Most prominently, factors reported to be engaged in early stages of acute viral infection were affected, e.g. entry, integration and provirus transcription and other cellular responses such as apoptosis and proliferation were modulated. For experimental validation of the gene expression analyses and computational predictions, individual pathways/sub-networks and significantly altered key regulators were investigated further. We showed that the expression of caveolin-1 (Cav-1), the top hub in the affected protein-protein interaction network, was significantly upregulated in chronically SIV-infected CD4+ T cells. Cav-1 is the main determinant of caveolae and a central component of several signal transduction pathways. Furthermore, CD4 downregulation and modulation of the expression of alternate and co-receptors as well as pathways associated with viral integration into the genome were also observed in these cells. Putatively, these modifications interfere with re-infection and the early replication cycle and inhibit cell death provoked by syncytia formation and bystander apoptosis. Conclusions Thus, by using the novel approach for network analysis, ICF, we predict that in the T cell line chronically infected with SIV, cellular processes that are known to be crucial for early phases of HIV/SIV replication are altered and cellular responses that result in cell death are modulated. These modifications presumably contribute to cell survival despite chronic infection.
Identification of molecular sub-networks associated with cell survival in a chronically SIVmac-infected human CD4+ T cell line
Journal Article
en
He et al.; licensee BioMed Central Ltd.
2018-06-13T00:40:48Z
Abstract
Background
The deciphering of cellular networks to determine susceptibility to infection by HIV or the related simian immunodeficiency virus (SIV) is a major challenge in infection biology.
Results
Here, we have compared gene expression profiles of a human CD4+ T cell line at 24 h after infection with a cell line of the same origin permanently releasing SIVmac. A new knowledge-based-network approach (Inter-Chain-Finder, ICF) has been used to identify sub-networks associated with cell survival of a chronically SIV-infected T cell line. Notably, the method can identify not only differentially expressed key hub genes but also non-differentially expressed, critical, ‘hidden’ regulators. Six out of the 13 predicted major hidden key regulators were among the landscape of proteins known to interact with HIV. Several sub-networks were dysregulated upon chronic infection with SIV. Most prominently, factors reported to be engaged in early stages of acute viral infection were affected, e.g. entry, integration and provirus transcription and other cellular responses such as apoptosis and proliferation were modulated. For experimental validation of the gene expression analyses and computational predictions, individual pathways/sub-networks and significantly altered key regulators were investigated further. We showed that the expression of caveolin-1 (Cav-1), the top hub in the affected protein-protein interaction network, was significantly upregulated in chronically SIV-infected CD4+ T cells. Cav-1 is the main determinant of caveolae and a central component of several signal transduction pathways. Furthermore, CD4 downregulation and modulation of the expression of alternate and co-receptors as well as pathways associated with viral integration into the genome were also observed in these cells. Putatively, these modifications interfere with re-infection and the early replication cycle and inhibit cell death provoked by syncytia formation and bystander apoptosis.
Conclusions
Thus, by using the novel approach for network analysis, ICF, we predict that in the T cell line chronically infected with SIV, cellular processes that are known to be crucial for early phases of HIV/SIV replication are altered and cellular responses that result in cell death are modulated. These modifications presumably contribute to cell survival despite chronic infection.
THUMBNAIL
12985_2013_Article_2486.pdf.jpg
12985_2013_Article_2486.pdf.jpg
Generated Thumbnail
image/jpeg
99296
https://hzi.openrepository.com/bitstream/10033/620783/5/12985_2013_Article_2486.pdf.jpg
63a94ea325cfab6ec9570e2f33829fc7
MD5
5
false
TEXT
12985_2013_Article_2486.pdf.txt
12985_2013_Article_2486.pdf.txt
Extracted Text
text/plain
113394
https://hzi.openrepository.com/bitstream/10033/620783/4/12985_2013_Article_2486.pdf.txt
645360ade29f0bbb7c69f1de6abd1851
MD5
4
false
ORIGINAL
12985_2013_Article_2486.pdf
application/pdf
2339429
https://hzi.openrepository.com/bitstream/10033/620783/1/12985_2013_Article_2486.pdf
e5cec4816ecd99bd7715d0459a5207c6
MD5
1
true
LICENSE
license.txt
license.txt
text/plain
1685
https://hzi.openrepository.com/bitstream/10033/620783/2/license.txt
cb598eeb10bfed09d26fd8d285172ad4
MD5
2
false
SWORD
art_122623938197306435.zip
SWORD deposit package
application/octet-stream
2045279
https://hzi.openrepository.com/bitstream/10033/620783/3/art_122623938197306435.zip
681d3c2dfda0629c4b79b3285c682bfd
MD5
3
false
10033/620783
oai:hzi.openrepository.com:10033/620783
2018-06-13 00:40:48.423
Helmholtz Zentrum für Infektionsforschung Repository
hzi@openrepository.com
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
oai:repository.helmholtz-hzi.de:10033/6206042019-08-30T11:31:23Zcom_10033_311308col_10033_620561
Moes, Lorin
0e98042c6b951f0539c7a6ef35a9c215
500
Wirth, Manfred
fe784cba1512cc6541c18747f2cae0a0
500
2016-11-29T08:51:06Z
2016-11-29T08:51:06Z
2007-11-22
The internal initiation of translation in bovine viral diarrhea virus RNA depends on the presence of an RNA pseudoknot upstream of the initiation codon. 2007, 4:124 Virol. J.
1743-422X
18034871
10.1186/1743-422X-4-124
http://hdl.handle.net/10033/620604
Virology journal
Bovine viral diarrhea virus (BVDV) is the prototype representative of the pestivirus genus in the Flaviviridae family. It has been shown that the initiation of translation of BVDV RNA occurs by an internal ribosome entry mechanism mediated by the 5' untranslated region of the viral RNA 1. The 5' and 3' boundaries of the IRES of the cytopathic BVDV NADL have been mapped and it has been suggested that the IRES extends into the coding of the BVDV polyprotein 2. A putative pseudoknot structure has been recognized in the BVDV 5'UTR in close proximity to the AUG start codon. A pseudoknot structure is characteristic for flavivirus IRESes and in the case of the closely related classical swine fever virus (CSFV) and the more distantly related Hepatitis C virus (HCV) pseudoknot function in translation has been demonstrated.
To characterize the BVDV IRESes in detail, we studied the BVDV translational initiation by transfection of dicistronic expression plasmids into mammalian cells. A region coding for the amino terminus of the BVDV SD-1 polyprotein contributes considerably to efficient initiation of translation. The translation efficiency mediated by the IRES of BVDV strains NADL and SD-1 approximates the poliovirus type I IRES directed translation in BHK cells. Compared to the poliovirus IRES increased expression levels are mediated by the BVDV IRES of strain SD-1 in murine cell lines, while lower levels are observed in human cell lines. Site directed mutagenesis revealed that a RNA pseudoknot upstream of the initiator AUG is an important structural element for IRES function. Mutants with impaired ability to base pair in stem I or II lost their translational activity. In mutants with repaired base pairing either in stem 1 or in stem 2 full translational activity was restored. Thus, the BVDV IRES translation is dependent on the pseudoknot integrity. These features of the pestivirus IRES are reminiscent of those of the classical swine fever virus, a pestivirus, and the hepatitis C viruses, another genus of the Flaviviridae.
The IRES of the non-cytopathic BVDV SD-1 strain displays features known from other pestivirus IRESes. The predicted pseudoknot in the 5'UTR of BVDV SD-1 virus represents an important structural element in BVDV translation.
ENG
Archived with thanks to Virology journal
5' Untranslated Regions
Animals
Cattle
Cell Line
Codon, Initiator
Diarrhea Viruses, Bovine Viral
HeLa Cells
Humans
Nucleic Acid Conformation
Protein Biosynthesis
RNA, Viral
Regulatory Sequences, Nucleic Acid
Structure-Activity Relationship
Transcription, Genetic
The internal initiation of translation in bovine viral diarrhea virus RNA depends on the presence of an RNA pseudoknot upstream of the initiation codon.
2018-06-13T02:26:29Z
Bovine viral diarrhea virus (BVDV) is the prototype representative of the pestivirus genus in the Flaviviridae family. It has been shown that the initiation of translation of BVDV RNA occurs by an internal ribosome entry mechanism mediated by the 5' untranslated region of the viral RNA 1. The 5' and 3' boundaries of the IRES of the cytopathic BVDV NADL have been mapped and it has been suggested that the IRES extends into the coding of the BVDV polyprotein 2. A putative pseudoknot structure has been recognized in the BVDV 5'UTR in close proximity to the AUG start codon. A pseudoknot structure is characteristic for flavivirus IRESes and in the case of the closely related classical swine fever virus (CSFV) and the more distantly related Hepatitis C virus (HCV) pseudoknot function in translation has been demonstrated.
To characterize the BVDV IRESes in detail, we studied the BVDV translational initiation by transfection of dicistronic expression plasmids into mammalian cells. A region coding for the amino terminus of the BVDV SD-1 polyprotein contributes considerably to efficient initiation of translation. The translation efficiency mediated by the IRES of BVDV strains NADL and SD-1 approximates the poliovirus type I IRES directed translation in BHK cells. Compared to the poliovirus IRES increased expression levels are mediated by the BVDV IRES of strain SD-1 in murine cell lines, while lower levels are observed in human cell lines. Site directed mutagenesis revealed that a RNA pseudoknot upstream of the initiator AUG is an important structural element for IRES function. Mutants with impaired ability to base pair in stem I or II lost their translational activity. In mutants with repaired base pairing either in stem 1 or in stem 2 full translational activity was restored. Thus, the BVDV IRES translation is dependent on the pseudoknot integrity. These features of the pestivirus IRES are reminiscent of those of the classical swine fever virus, a pestivirus, and the hepatitis C viruses, another genus of the Flaviviridae.
The IRES of the non-cytopathic BVDV SD-1 strain displays features known from other pestivirus IRESes. The predicted pseudoknot in the 5'UTR of BVDV SD-1 virus represents an important structural element in BVDV translation.
ORIGINAL
12985_2007_Article_337.pdf
Open Access publication
application/pdf
391671
https://hzi.openrepository.com/bitstream/10033/620604/1/12985_2007_Article_337.pdf
5c190612568a17aa94c640a17ed6253a
MD5
1
true
LICENSE
license.txt
license.txt
text/plain
1685
https://hzi.openrepository.com/bitstream/10033/620604/2/license.txt
cb598eeb10bfed09d26fd8d285172ad4
MD5
2
false
SWORD
art_766591435122287968.zip
SWORD deposit package
application/octet-stream
288008
https://hzi.openrepository.com/bitstream/10033/620604/3/art_766591435122287968.zip
030da88ef4e1f6e1929d94a7f0a867e2
MD5
3
false
TEXT
12985_2007_Article_337.pdf.txt
12985_2007_Article_337.pdf.txt
Extracted Text
text/plain
64345
https://hzi.openrepository.com/bitstream/10033/620604/4/12985_2007_Article_337.pdf.txt
b51b479fce29254139501afc71d08759
MD5
4
false
THUMBNAIL
12985_2007_Article_337.pdf.jpg
12985_2007_Article_337.pdf.jpg
Generated Thumbnail
image/jpeg
86296
https://hzi.openrepository.com/bitstream/10033/620604/5/12985_2007_Article_337.pdf.jpg
da0c791fecc1b70170cc8973cf9ad052
MD5
5
false
10033/620604
oai:hzi.openrepository.com:10033/620604
2019-08-30 11:31:23.518
Helmholtz Zentrum für Infektionsforschung Repository
hzi@openrepository.com
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
oai:repository.helmholtz-hzi.de:10033/6206882019-08-30T11:33:57Zcom_10033_311308com_10033_620618col_10033_620619col_10033_559591col_10033_620561
Bock, Tobias
18684ebad3d88945a6982360946fa17c
500
Volz, Carsten
e8b23b910d325e5e91a028a1b34f39d1
500
Hering, Vanessa
16bfdc35fc4a470c30c9578ef63257b3
500
Scrima, Andrea
c3ad854d78113b599fa24cb5e9e6fbcc
500
Müller, Rolf
38d0f8dc80ae572919b57a1e4d8baf29
500
Blankenfeldt, Wulf
2e008ccc762d1395649412fb11521159
600
http://orcid.org/0000-0001-9886-9668
Hel,holtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.
2017-01-09T15:31:16Z
2017-01-09T15:31:16Z
2016-12-09
The AibR-isovaleryl coenzyme A regulator and its DNA binding site - a model for the regulation of alternative de novo isovaleryl coenzyme A biosynthesis in Myxococcus xanthus. 2016 Nucleic Acids Res.
1362-4962
27940564
10.1093/nar/gkw1238
http://hdl.handle.net/10033/620688
Nucleic acids research
Isovaleryl coenzyme A (IV-CoA) is an important building block of iso-fatty acids. In myxobacteria, IV-CoA is essential for the formation of signaling molecules involved in fruiting body formation. Leucine degradation is the common source of IV-CoA, but a second, de novo biosynthetic route to IV-CoA termed AIB (alternative IV-CoA biosynthesis) was recently discovered in M. xanthus The AIB-operon contains the TetR-like transcriptional regulator AibR, which we characterize in this study. We demonstrate that IV-CoA binds AibR with micromolar affinity and show by gelshift experiments that AibR interacts with the promoter region of the AIB-operon once IV-CoA is present. We identify an 18-bp near-perfect palindromic repeat as containing the AibR operator and provide evidence that AibR also controls an additional genomic locus coding for a putative acetyl-CoA acetyltransferase. To elucidate atomic details, we determined crystal structures of AibR in the apo, the IV-CoA- and the IV-CoA-DNA-bound state to 1.7 Å, 2.35 Å and 2.92 Å, respectively. IV-CoA induces partial unfolding of an α-helix, which allows sequence-specific interactions between AibR and its operator. This study provides insights into AibR-mediated regulation and shows that AibR functions in an unusual TetR-like manner by blocking transcription not in the ligand-free but in the effector-bound state.
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
The AibR-isovaleryl coenzyme A regulator and its DNA binding site - a model for the regulation of alternative de novo isovaleryl coenzyme A biosynthesis in Myxococcus xanthus.
Article
2018-06-12T20:00:59Z
Isovaleryl coenzyme A (IV-CoA) is an important building block of iso-fatty acids. In myxobacteria, IV-CoA is essential for the formation of signaling molecules involved in fruiting body formation. Leucine degradation is the common source of IV-CoA, but a second, de novo biosynthetic route to IV-CoA termed AIB (alternative IV-CoA biosynthesis) was recently discovered in M. xanthus The AIB-operon contains the TetR-like transcriptional regulator AibR, which we characterize in this study. We demonstrate that IV-CoA binds AibR with micromolar affinity and show by gelshift experiments that AibR interacts with the promoter region of the AIB-operon once IV-CoA is present. We identify an 18-bp near-perfect palindromic repeat as containing the AibR operator and provide evidence that AibR also controls an additional genomic locus coding for a putative acetyl-CoA acetyltransferase. To elucidate atomic details, we determined crystal structures of AibR in the apo, the IV-CoA- and the IV-CoA-DNA-bound state to 1.7 Å, 2.35 Å and 2.92 Å, respectively. IV-CoA induces partial unfolding of an α-helix, which allows sequence-specific interactions between AibR and its operator. This study provides insights into AibR-mediated regulation and shows that AibR functions in an unusual TetR-like manner by blocking transcription not in the ligand-free but in the effector-bound state.
ORIGINAL
Bock et al.pdf
Bock et al.pdf
Open Access publication
application/pdf
8077125
https://hzi.openrepository.com/bitstream/10033/620688/1/Bock%20et%20al.pdf
d72e6c954acd39217c0147b14657eb02
MD5
1
true
CC-LICENSE
license_url
license_url
text/plain
49
https://hzi.openrepository.com/bitstream/10033/620688/2/license_url
924993ce0b3ba389f79f32a1b2735415
MD5
2
false
license_text
license_text
application/octet-stream
0
https://hzi.openrepository.com/bitstream/10033/620688/3/license_text
d41d8cd98f00b204e9800998ecf8427e
MD5
3
false
license_rdf
license_rdf
application/octet-stream
0
https://hzi.openrepository.com/bitstream/10033/620688/4/license_rdf
d41d8cd98f00b204e9800998ecf8427e
MD5
4
false
LICENSE
license.txt
license.txt
text/plain
1685
https://hzi.openrepository.com/bitstream/10033/620688/5/license.txt
cb598eeb10bfed09d26fd8d285172ad4
MD5
5
false
TEXT
Bock et al.pdf.txt
Bock et al.pdf.txt
Extracted Text
text/plain
60943
https://hzi.openrepository.com/bitstream/10033/620688/6/Bock%20et%20al.pdf.txt
ae476376f7b2a75f2a419ed5dcbaadc7
MD5
6
false
THUMBNAIL
Bock et al.pdf.jpg
Bock et al.pdf.jpg
Generated Thumbnail
image/jpeg
183323
https://hzi.openrepository.com/bitstream/10033/620688/7/Bock%20et%20al.pdf.jpg
53cc33d6bc099d71646352408f206d4e
MD5
7
false
10033/620688
oai:hzi.openrepository.com:10033/620688
2019-08-30 11:33:57.486
Helmholtz Zentrum für Infektionsforschung Repository
hzi@openrepository.com
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
oai:repository.helmholtz-hzi.de:10033/6211002021-07-05T15:12:57Zcom_10033_311308com_10033_128109com_10033_622921col_10033_622922col_10033_621829col_10033_620561
Chaudhry, M Zeeshan
e86c0c4d7c8e9bd86b0c1c523ac2f28a
500
Kasmapour, Bahram
9241b10872d1fdbfc72851662f76b489
500
Plaza-Sirvent, Carlos
39ece84a3ce6830bb74c00e89f1d5c44
500
Bajagic, Milica
3d3b97feffdeb501621193c0b09cd441
500
Casalegno Garduño, Rosaely
7294668a05de21d73e74e3e658a1b5b4
500
Borkner, Lisa
b6d2c71ea1f0417aa71db7693d78b3b9
500
Lenac Roviš, Tihana
e0ef198a965cef3e88aabc48d003d37b
500
Scrima, Andrea
c3ad854d78113b599fa24cb5e9e6fbcc
500
Jonjic, Stipan
200f21c036e9f0503976ae2527331ed8
500
Schmitz, Ingo
d064a965762a0bd3539b3090354dba25
600
http://orcid.org/0000-0002-5360-0419
Cicin-Sain, Luka
e159d8d92d15f2bb5406d303ade66cbb
600
http://orcid.org/0000-0003-3978-778X
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
2017-09-11T10:30:46Z
2017-09-11T10:30:46Z
2017
UL36 Rescues Apoptosis Inhibition and In vivo Replication of a Chimeric MCMV Lacking the M36 Gene. 2017, 7:312 Front Cell Infect Microbiol
2235-2988
28770171
10.3389/fcimb.2017.00312
http://hdl.handle.net/10033/621100
Frontiers in cellular and infection microbiology
Apoptosis is an important defense mechanism mounted by the immune system to control virus replication. Hence, cytomegaloviruses (CMV) evolved and acquired numerous anti-apoptotic genes. The product of the human CMV (HCMV) UL36 gene, pUL36 (also known as vICA), binds to pro-caspase-8, thus inhibiting death-receptor apoptosis and enabling viral replication in differentiated THP-1 cells. In vivo studies of the function of HCMV genes are severely limited due to the strict host specificity of cytomegaloviruses, but CMV orthologues that co-evolved with other species allow the experimental study of CMV biology in vivo. The mouse CMV (MCMV) homolog of the UL36 gene is called M36, and its protein product (pM36) is a functional homolog of vICA that binds to murine caspase-8 and inhibits its activation. M36-deficient MCMV is severely growth impaired in macrophages and in vivo. Here we show that pUL36 binds to the murine pro-caspase-8, and that UL36 expression inhibits death-receptor apoptosis in murine cells and can replace M36 to allow MCMV growth in vitro and in vivo. We generated a chimeric MCMV expressing the UL36 ORF sequence instead of the M36 one. The newly generated MCMV(UL36) inhibited apoptosis in macrophage lines RAW 264.7, J774A.1, and IC-21 and its growth was rescued to wild type levels. Similarly, growth was rescued in vivo in the liver and spleen, but only partially in the salivary glands of BALB/c and C57BL/6 mice. In conclusion, we determined that an immune-evasive HCMV gene is conserved enough to functionally replace its MCMV counterpart and thus allow its study in an in vivo setting. As UL36 and M36 proteins engage the same molecular host target, our newly developed model can facilitate studies of anti-viral compounds targeting pUL36 in vivo.
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
UL36 Rescues Apoptosis Inhibition and In vivo Replication of a Chimeric MCMV Lacking the M36 Gene.
Article
2018-06-12T22:18:39Z
Apoptosis is an important defense mechanism mounted by the immune system to control virus replication. Hence, cytomegaloviruses (CMV) evolved and acquired numerous anti-apoptotic genes. The product of the human CMV (HCMV) UL36 gene, pUL36 (also known as vICA), binds to pro-caspase-8, thus inhibiting death-receptor apoptosis and enabling viral replication in differentiated THP-1 cells. In vivo studies of the function of HCMV genes are severely limited due to the strict host specificity of cytomegaloviruses, but CMV orthologues that co-evolved with other species allow the experimental study of CMV biology in vivo. The mouse CMV (MCMV) homolog of the UL36 gene is called M36, and its protein product (pM36) is a functional homolog of vICA that binds to murine caspase-8 and inhibits its activation. M36-deficient MCMV is severely growth impaired in macrophages and in vivo. Here we show that pUL36 binds to the murine pro-caspase-8, and that UL36 expression inhibits death-receptor apoptosis in murine cells and can replace M36 to allow MCMV growth in vitro and in vivo. We generated a chimeric MCMV expressing the UL36 ORF sequence instead of the M36 one. The newly generated MCMV(UL36) inhibited apoptosis in macrophage lines RAW 264.7, J774A.1, and IC-21 and its growth was rescued to wild type levels. Similarly, growth was rescued in vivo in the liver and spleen, but only partially in the salivary glands of BALB/c and C57BL/6 mice. In conclusion, we determined that an immune-evasive HCMV gene is conserved enough to functionally replace its MCMV counterpart and thus allow its study in an in vivo setting. As UL36 and M36 proteins engage the same molecular host target, our newly developed model can facilitate studies of anti-viral compounds targeting pUL36 in vivo.
ORIGINAL
Chaudhry et al.pdf
Chaudhry et al.pdf
Open Access publication
application/pdf
2196057
https://repository.helmholtz-hzi.de/bitstream/10033/621100/1/Chaudhry%20et%20al.pdf
f45d204a947de92d36ac55a5f4f2f70e
MD5
1
true
supplementary figure 1.pdf
supplementary figure 1.pdf
supplementary figure 1
application/pdf
128533
https://repository.helmholtz-hzi.de/bitstream/10033/621100/6/supplementary%20figure%201.pdf
8b5262db02a4458c1e1a7b50539d380b
MD5
6
false
CC-LICENSE
license_url
license_url
text/plain
49
https://repository.helmholtz-hzi.de/bitstream/10033/621100/2/license_url
924993ce0b3ba389f79f32a1b2735415
MD5
2
false
license_text
license_text
application/octet-stream
0
https://repository.helmholtz-hzi.de/bitstream/10033/621100/3/license_text
d41d8cd98f00b204e9800998ecf8427e
MD5
3
false
license_rdf
license_rdf
application/octet-stream
0
https://repository.helmholtz-hzi.de/bitstream/10033/621100/4/license_rdf
d41d8cd98f00b204e9800998ecf8427e
MD5
4
false
LICENSE
license.txt
license.txt
text/plain
1685
https://repository.helmholtz-hzi.de/bitstream/10033/621100/5/license.txt
cb598eeb10bfed09d26fd8d285172ad4
MD5
5
false
TEXT
Chaudhry et al.pdf.txt
Chaudhry et al.pdf.txt
Extracted Text
text/plain
54034
https://repository.helmholtz-hzi.de/bitstream/10033/621100/7/Chaudhry%20et%20al.pdf.txt
8af4ee1e0d274ab076434a6e670d43ae
MD5
7
false
supplementary figure 1.pdf.txt
supplementary figure 1.pdf.txt
Extracted Text
text/plain
525
https://repository.helmholtz-hzi.de/bitstream/10033/621100/9/supplementary%20figure%201.pdf.txt
3e9345ca36a33f60b8bd7986822f9b07
MD5
9
false
THUMBNAIL
Chaudhry et al.pdf.jpg
Chaudhry et al.pdf.jpg
Generated Thumbnail
image/jpeg
120378
https://repository.helmholtz-hzi.de/bitstream/10033/621100/11/Chaudhry%20et%20al.pdf.jpg
0db77f044c39f2fdf0b65510ea2a28bc
MD5
11
false
supplementary figure 1.pdf.jpg
supplementary figure 1.pdf.jpg
Generated Thumbnail
image/jpeg
36503
https://repository.helmholtz-hzi.de/bitstream/10033/621100/12/supplementary%20figure%201.pdf.jpg
4c2b05c48436097e0023939eb729567a
MD5
12
false
10033/621100
oai:repository.helmholtz-hzi.de:10033/621100
2021-07-05 15:12:57.751
Helmholtz Zentrum für Infektionsforschung Repository
hzi@openrepository.com
Tk9OLUVYQ0xVU0lWRSBESVNUUklCVVRJT04gTElDRU5TRQoKQnkgc2lnbmluZyBhbmQgc3VibWl0dGluZyB0aGlzIGxpY2Vuc2UsIHlvdSAodGhlIGF1dGhvcihzKSBvciBjb3B5cmlnaHQKb3duZXIpIGdyYW50cyB0byBIZWxtaG9sdHogWmVudHJ1bSBm77+9ciBJbmZla3Rpb25zZm9yc2NodW5nIFJlcG9zaXRvcnkgKEhaSSkgdGhlIG5vbi1leGNsdXNpdmUgcmlnaHQgdG8gcmVwcm9kdWNlLAp0cmFuc2xhdGUgKGFzIGRlZmluZWQgYmVsb3cpLCBhbmQvb3IgZGlzdHJpYnV0ZSB5b3VyIHN1Ym1pc3Npb24gKGluY2x1ZGluZwp0aGUgYWJzdHJhY3QpIHdvcmxkd2lkZSBpbiBwcmludCBhbmQgZWxlY3Ryb25pYyBmb3JtYXQgYW5kIGluIGFueSBtZWRpdW0sCmluY2x1ZGluZyBidXQgbm90IGxpbWl0ZWQgdG8gYXVkaW8gb3IgdmlkZW8uCgpZb3UgYWdyZWUgdGhhdCBIWkkgbWF5LCB3aXRob3V0IGNoYW5naW5nIHRoZSBjb250ZW50LCB0cmFuc2xhdGUgdGhlCnN1Ym1pc3Npb24gdG8gYW55IG1lZGl1bSBvciBmb3JtYXQgZm9yIHRoZSBwdXJwb3NlIG9mIHByZXNlcnZhdGlvbi4KCllvdSBhbHNvIGFncmVlIHRoYXQgSFpJIG1heSBrZWVwIG1vcmUgdGhhbiBvbmUgY29weSBvZiB0aGlzIHN1Ym1pc3Npb24gZm9yCnB1cnBvc2VzIG9mIHNlY3VyaXR5LCBiYWNrLXVwIGFuZCBwcmVzZXJ2YXRpb24uCgpZb3UgcmVwcmVzZW50IHRoYXQgdGhlIHN1Ym1pc3Npb24gaXMgeW91ciBvcmlnaW5hbCB3b3JrLCBhbmQgdGhhdCB5b3UgaGF2ZQp0aGUgcmlnaHQgdG8gZ3JhbnQgdGhlIHJpZ2h0cyBjb250YWluZWQgaW4gdGhpcyBsaWNlbnNlLiBZb3UgYWxzbyByZXByZXNlbnQKdGhhdCB5b3VyIHN1Ym1pc3Npb24gZG9lcyBub3QsIHRvIHRoZSBiZXN0IG9mIHlvdXIga25vd2xlZGdlLCBpbmZyaW5nZSB1cG9uCmFueW9uZSdzIGNvcHlyaWdodC4KCklmIHRoZSBzdWJtaXNzaW9uIGNvbnRhaW5zIG1hdGVyaWFsIGZvciB3aGljaCB5b3UgZG8gbm90IGhvbGQgY29weXJpZ2h0LAp5b3UgcmVwcmVzZW50IHRoYXQgeW91IGhhdmUgb2J0YWluZWQgdGhlIHVucmVzdHJpY3RlZCBwZXJtaXNzaW9uIG9mIHRoZQpjb3B5cmlnaHQgb3duZXIgdG8gZ3JhbnQgSFpJIHRoZSByaWdodHMgcmVxdWlyZWQgYnkgdGhpcyBsaWNlbnNlLCBhbmQgdGhhdApzdWNoIHRoaXJkLXBhcnR5IG93bmVkIG1hdGVyaWFsIGlzIGNsZWFybHkgaWRlbnRpZmllZCBhbmQgYWNrbm93bGVkZ2VkCndpdGhpbiB0aGUgdGV4dCBvciBjb250ZW50IG9mIHRoZSBzdWJtaXNzaW9uLgoKSUYgVEhFIFNVQk1JU1NJT04gSVMgQkFTRUQgVVBPTiBXT1JLIFRIQVQgSEFTIEJFRU4gU1BPTlNPUkVEIE9SIFNVUFBPUlRFRApCWSBBTiBBR0VOQ1kgT1IgT1JHQU5JWkFUSU9OIE9USEVSIFRIQU4gSFpJLCBZT1UgUkVQUkVTRU5UIFRIQVQgWU9VIEhBVkUKRlVMRklMTEVEIEFOWSBSSUdIVCBPRiBSRVZJRVcgT1IgT1RIRVIgT0JMSUdBVElPTlMgUkVRVUlSRUQgQlkgU1VDSApDT05UUkFDVCBPUiBBR1JFRU1FTlQuCgpIWkkgd2lsbCBjbGVhcmx5IGlkZW50aWZ5IHlvdXIgbmFtZShzKSBhcyB0aGUgYXV0aG9yKHMpIG9yIG93bmVyKHMpIG9mIHRoZQpzdWJtaXNzaW9uLCBhbmQgd2lsbCBub3QgbWFrZSBhbnkgYWx0ZXJhdGlvbiwgb3RoZXIgdGhhbiBhcyBhbGxvd2VkIGJ5IHRoaXMKbGljZW5zZSwgdG8geW91ciBzdWJtaXNzaW9uLgo=
oai:repository.helmholtz-hzi.de:10033/6211572019-08-30T11:27:16Zcom_10033_311308col_10033_620561
Archna, Archna
ac5e6e9ac4374726c8cd22fa38487c20
500
Scrima, Andrea
c3ad854d78113b599fa24cb5e9e6fbcc
500
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr.7, 38124 Braunschweig, Germany.
2017-11-03T10:13:04Z
2017-11-03T10:13:04Z
2017-10-01
Identification, biochemical characterization and crystallization of the central region of human ATG16L1. 2017, 73 (Pt 10):560-567 Acta Crystallogr F Struct Biol Commun
2053-230X
28994404
10.1107/S2053230X17013280
http://hdl.handle.net/10033/621157
Acta crystallographica. Section F, Structural biology communications
ATG16L1 plays a major role in autophagy. It acts as a molecular scaffold which mediates protein-protein interactions essential for autophagosome formation. The ATG12~ATG5-ATG16L1 complex is one of the key complexes involved in autophagosome formation. Human ATG16L1 comprises 607 amino acids with three functional domains named ATG5BD, CCD and WD40, where the C-terminal WD40 domain represents approximately 50% of the full-length protein. Previously, structures of the C-terminal WD40 domain of human ATG16L1 as well as of human ATG12~ATG5 in complex with the ATG5BD of ATG16L1 have been reported. However, apart from the ATG5BD, no structural information for the N-terminal half, including the CCD, of human ATG16L1 is available. In this study, the authors aimed to structurally characterize the N-terminal half of ATG16L1. ATG16L111-307 in complex with ATG5 has been purified and crystallized in two crystal forms. However, both crystal structures revealed degradation of ATG16L1, resulting in crystals comprising only full-length ATG5 and the ATG5BD of ATG16L1. The structures of ATG5-ATG5BD in two novel crystal forms are presented, further supporting the previously observed dimerization of ATG5-ATG16L1. The reported degradation points towards a high instability at the linker region between the ATG5BD and the CCD in ATG16L1. Based on this observation and further biochemical analysis of ATG16L1, a stable 236-amino-acid subfragment comprising residues 72-307 of the N-terminal half of ATG16L1, covering the residual, so far structurally uncharacterized region of human ATG16L1, was identified. Here, the identification, purification, biochemical characterization and crystallization of the proteolytically stable ATG16L172-307 subfragment are reported.
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
Identification, biochemical characterization and crystallization of the central region of human ATG16L1.
Article
2018-06-13T19:46:28Z
ATG16L1 plays a major role in autophagy. It acts as a molecular scaffold which mediates protein-protein interactions essential for autophagosome formation. The ATG12~ATG5-ATG16L1 complex is one of the key complexes involved in autophagosome formation. Human ATG16L1 comprises 607 amino acids with three functional domains named ATG5BD, CCD and WD40, where the C-terminal WD40 domain represents approximately 50% of the full-length protein. Previously, structures of the C-terminal WD40 domain of human ATG16L1 as well as of human ATG12~ATG5 in complex with the ATG5BD of ATG16L1 have been reported. However, apart from the ATG5BD, no structural information for the N-terminal half, including the CCD, of human ATG16L1 is available. In this study, the authors aimed to structurally characterize the N-terminal half of ATG16L1. ATG16L111-307 in complex with ATG5 has been purified and crystallized in two crystal forms. However, both crystal structures revealed degradation of ATG16L1, resulting in crystals comprising only full-length ATG5 and the ATG5BD of ATG16L1. The structures of ATG5-ATG5BD in two novel crystal forms are presented, further supporting the previously observed dimerization of ATG5-ATG16L1. The reported degradation points towards a high instability at the linker region between the ATG5BD and the CCD in ATG16L1. Based on this observation and further biochemical analysis of ATG16L1, a stable 236-amino-acid subfragment comprising residues 72-307 of the N-terminal half of ATG16L1, covering the residual, so far structurally uncharacterized region of human ATG16L1, was identified. Here, the identification, purification, biochemical characterization and crystallization of the proteolytically stable ATG16L172-307 subfragment are reported.
ORIGINAL
Archna and Scrima.pdf
Archna and Scrima.pdf
allowed publisher's PDF
application/pdf
1181250
https://hzi.openrepository.com/bitstream/10033/621157/1/Archna%20and%20Scrima.pdf
c0a732162a45348175d48f81bc2a0af1
MD5
1
true
supporting figures and tables.pdf
supporting figures and tables.pdf
supporting figures and tables
application/pdf
1078558
https://hzi.openrepository.com/bitstream/10033/621157/2/supporting%20figures%20and%20tables.pdf
36c46336d7e9545e7ab3681c21374956
MD5
2
false
CC-LICENSE
license_url
license_url
text/plain
49
https://hzi.openrepository.com/bitstream/10033/621157/3/license_url
924993ce0b3ba389f79f32a1b2735415
MD5
3
false
license_text
license_text
application/octet-stream
0
https://hzi.openrepository.com/bitstream/10033/621157/4/license_text
d41d8cd98f00b204e9800998ecf8427e
MD5
4
false
license_rdf
license_rdf
application/octet-stream
0
https://hzi.openrepository.com/bitstream/10033/621157/5/license_rdf
d41d8cd98f00b204e9800998ecf8427e
MD5
5
false
LICENSE
license.txt
license.txt
text/plain
1685
https://hzi.openrepository.com/bitstream/10033/621157/6/license.txt
cb598eeb10bfed09d26fd8d285172ad4
MD5
6
false
TEXT
Archna and Scrima.pdf.txt
Archna and Scrima.pdf.txt
Extracted Text
text/plain
29806
https://hzi.openrepository.com/bitstream/10033/621157/7/Archna%20and%20Scrima.pdf.txt
f93d36640333b1e17738a41810f9056e
MD5
7
false
supporting figures and tables.pdf.txt
supporting figures and tables.pdf.txt
Extracted Text
text/plain
5990
https://hzi.openrepository.com/bitstream/10033/621157/9/supporting%20figures%20and%20tables.pdf.txt
900bf43e759518709c1c5e7483e60aa3
MD5
9
false
THUMBNAIL
Archna and Scrima.pdf.jpg
Archna and Scrima.pdf.jpg
Generated Thumbnail
image/jpeg
111791
https://hzi.openrepository.com/bitstream/10033/621157/11/Archna%20and%20Scrima.pdf.jpg
df6316c3356f949e1bff28db77a55fb9
MD5
11
false
supporting figures and tables.pdf.jpg
supporting figures and tables.pdf.jpg
Generated Thumbnail
image/jpeg
19771
https://hzi.openrepository.com/bitstream/10033/621157/12/supporting%20figures%20and%20tables.pdf.jpg
2f18c2d83598c8e06c8ffc7508705e2c
MD5
12
false
10033/621157
oai:hzi.openrepository.com:10033/621157
2019-08-30 11:27:16.144
Helmholtz Zentrum für Infektionsforschung Repository
hzi@openrepository.com
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
oai:repository.helmholtz-hzi.de:10033/6213212019-08-30T11:29:17Zcom_10033_311308col_10033_620561
Fiebig, David
20b139c4c62dc04af4e92f65da292fce
500
Schmelz, Stefan
91c14d4d519c8fc0f1446b2262626ea5
500
Zindel, Stephan
a61c772c34490987d20ae8f7d9270b48
500
Ehret, Vera
0b5e1149b74b5831d421597cb02f189c
500
Beck, Jan
022b7af2d2e5908021fd79137745e775
500
Ebenig, Aileen
52c2e65550ba13d57a921c47034f7574
500
Ehret, Marina
dfbef4acccc606332c5564dd53eb3dc1
500
Fröls, Sabrina
9b104580758cd0563af2cdc2fa9d108b
500
Pfeifer, Felicitas
d9c60d4d313b8ca23418cedfe1b9d133
500
Kolmar, Harald
5989991fb885a1d13b170f2afce3dcce
500
Fuchsbauer, Hans-Lothar
be4a81b2a16b231e991ac87fffdc1f69
500
Scrima, Andrea
c3ad854d78113b599fa24cb5e9e6fbcc
500
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
2018-03-12T15:28:28Z
2018-03-12T15:28:28Z
2016
Structure of the Dispase Autolysis-inducing Protein from Streptomyces mobaraensis and Glutamine Cross-linking Sites for Transglutaminase. 2016, 291 (39):20417-26 J. Biol. Chem.
1083-351X
27493205
10.1074/jbc.M116.731109
http://hdl.handle.net/10033/621321
The Journal of biological chemistry
PMC5034039
Transglutaminase from Streptomyces mobaraensis (MTG) is an important enzyme for cross-linking and modifying proteins. An intrinsic substrate of MTG is the dispase autolysis-inducing protein (DAIP). The amino acid sequence of DAIP contains 5 potential glutamines and 10 lysines for MTG-mediated cross-linking. The aim of the study was to determine the structure and glutamine cross-linking sites of the first physiological MTG substrate. A production procedure was established in Escherichia coli BL21 (DE3) to obtain high yields of recombinant DAIP. DAIP variants were prepared by replacing four of five glutamines for asparagines in various combinations via site-directed mutagenesis. Incorporation of biotin cadaverine revealed a preference of MTG for the DAIP glutamines in the order of Gln-39 ≫ Gln-298 > Gln-345 ∼ Gln-65 ≫ Gln-144. In the structure of DAIP the preferred glutamines do cluster at the top of the seven-bladed β-propeller. This suggests a targeted cross-linking of DAIP by MTG that may occur after self-assembly in the bacterial cell wall. Based on our biochemical and structural data of the first physiological MTG substrate, we further provide novel insight into determinants of MTG-mediated modification, specificity, and efficiency.
en
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5034039/
http://creativecommons.org/licenses/by-nc-sa/4.0/
Bacterial Proteins
Escherichia coli
Recombinant Proteins
Streptomyces
Transglutaminases
Structure of the Dispase Autolysis-inducing Protein from Streptomyces mobaraensis and Glutamine Cross-linking Sites for Transglutaminase.
Article
2018-06-13T02:29:58Z
Transglutaminase from Streptomyces mobaraensis (MTG) is an important enzyme for cross-linking and modifying proteins. An intrinsic substrate of MTG is the dispase autolysis-inducing protein (DAIP). The amino acid sequence of DAIP contains 5 potential glutamines and 10 lysines for MTG-mediated cross-linking. The aim of the study was to determine the structure and glutamine cross-linking sites of the first physiological MTG substrate. A production procedure was established in Escherichia coli BL21 (DE3) to obtain high yields of recombinant DAIP. DAIP variants were prepared by replacing four of five glutamines for asparagines in various combinations via site-directed mutagenesis. Incorporation of biotin cadaverine revealed a preference of MTG for the DAIP glutamines in the order of Gln-39 ≫ Gln-298 > Gln-345 ∼ Gln-65 ≫ Gln-144. In the structure of DAIP the preferred glutamines do cluster at the top of the seven-bladed β-propeller. This suggests a targeted cross-linking of DAIP by MTG that may occur after self-assembly in the bacterial cell wall. Based on our biochemical and structural data of the first physiological MTG substrate, we further provide novel insight into determinants of MTG-mediated modification, specificity, and efficiency.
ORIGINAL
Fiebig et al.pdf
Fiebig et al.pdf
after embargo allowed publisher's PDF
application/pdf
2282977
https://hzi.openrepository.com/bitstream/10033/621321/1/Fiebig%20et%20al.pdf
0d8fb18e3b677d89c70fbd751068a013
MD5
1
true
CC-LICENSE
license_url
license_url
text/plain
49
https://hzi.openrepository.com/bitstream/10033/621321/2/license_url
924993ce0b3ba389f79f32a1b2735415
MD5
2
false
license_text
license_text
application/octet-stream
0
https://hzi.openrepository.com/bitstream/10033/621321/3/license_text
d41d8cd98f00b204e9800998ecf8427e
MD5
3
false
license_rdf
license_rdf
application/octet-stream
0
https://hzi.openrepository.com/bitstream/10033/621321/4/license_rdf
d41d8cd98f00b204e9800998ecf8427e
MD5
4
false
LICENSE
license.txt
license.txt
text/plain
1685
https://hzi.openrepository.com/bitstream/10033/621321/5/license.txt
cb598eeb10bfed09d26fd8d285172ad4
MD5
5
false
TEXT
Fiebig et al.pdf.txt
Fiebig et al.pdf.txt
Extracted Text
text/plain
53711
https://hzi.openrepository.com/bitstream/10033/621321/6/Fiebig%20et%20al.pdf.txt
64f3ceef92c55f6dd2b099e96c9bfe05
MD5
6
false
THUMBNAIL
Fiebig et al.pdf.jpg
Fiebig et al.pdf.jpg
Generated Thumbnail
image/jpeg
190758
https://hzi.openrepository.com/bitstream/10033/621321/7/Fiebig%20et%20al.pdf.jpg
82995e0ffaa8b633273cbc4e380717a4
MD5
7
false
10033/621321
oai:hzi.openrepository.com:10033/621321
2019-08-30 11:29:17.606
Helmholtz Zentrum für Infektionsforschung Repository
hzi@openrepository.com
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
oai:repository.helmholtz-hzi.de:10033/6213322019-08-30T11:29:17Zcom_10033_128109com_10033_620644com_10033_311308col_10033_128110col_10033_620646col_10033_620561
Sadana, Pooja
cf193c96d848ddcaa4d23ab201299a43
500
Geyer, Rebecca
e3ef70671d9a0201ffa2d9a378d79860
500
Pezoldt, Joern
12528b25f15cd5d3e26390b9840374ee
500
Helmsing, Saskia
ca7cb60d18b17a79c5e50da72d13b8e1
500
Huehn, Jochen
593aa066bacc8199a66ef7fe31379623
600
http://orcid.org/0000-0001-8071-1379
Hust, Michael
bd09260175734ab3f5987ca9fb155d94
500
Dersch, Petra
500f0a5abfe1d165918adfd28d1a4df3
600
http://orcid.org/0000-0001-8177-3280
Scrima, Andrea
c3ad854d78113b599fa24cb5e9e6fbcc
500
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
2018-03-23T15:04:02Z
2018-03-23T15:04:02Z
2018-03-13
The invasin D protein fromYersinia pseudotuberculosisselectively binds the Fab region of host antibodies and affects colonization of the intestine. 2018 J. Biol. Chem.
1083-351X
29535184
10.1074/jbc.RA117.001068
http://hdl.handle.net/10033/621332
The Journal of biological chemistry
Yersinia pseudotuberculosis is a Gram-negative bacterium and zoonotic pathogen responsible for a wide range of diseases, ranging from mild diarrhea, enterocolitis, lymphatic adenitis to persistent local inflammation. TheY. pseudotuberculosisinvasin D (InvD) molecule belongs to the invasin (InvA)-type autotransporter proteins, but its structure and function remain unknown. In this study, we present the first crystal structure of InvD, analyzed its expression and function in a murine infection model, and identified its target molecule in the host. We found that InvD is induced at 37°C and expressed in vivo2-4 days after infection, indicating that InvD is a virulence factor. During infection, InvD was expressed in all parts of the intestinal tract, but not in deeper lymphoid tissues. The crystal structure of the C-terminal adhesion domain of InvD revealed a distinct Ig-related fold, that, apart from the canonical β-sheets, comprises various modifications of and insertions into the Ig-core structure. We identified the Fab fragment of host-derived IgG/IgA antibodies as the target of the adhesion domain. Phage display panning and flow cytometry data further revealed that InvD exhibits a preferential binding specificity toward antibodies with VH3/VK1 variable domains and that it is specifically recruited to a subset of B cells. This finding suggests that InvD modulates Ig functions in the intestine and affects direct interactions with a subset of cell surface-exposed B-cell receptors. In summary, our results provide extensive insights into the structure of InvD and its specific interaction with the target molecule in the host.
en
http://creativecommons.org/licenses/by-nc-sa/4.0/
The invasin D protein fromYersinia pseudotuberculosisselectively binds the Fab region of host antibodies and affects colonization of the intestine.
Article
Yersinia pseudotuberculosis
is a Gram-negative bacterium and zoonotic pathogen responsible for a wide range of diseases, ranging from mild diarrhea, enterocolitis, lymphatic adenitis to persistent local inflammation. TheY. pseudotuberculosisinvasin D (InvD) molecule belongs to the invasin (InvA)-type autotransporter proteins, but its structure and function remain unknown. In this study, we present the first crystal structure of InvD, analyzed its expression and function in a murine infection model, and identified its target molecule in the host. We found that InvD is induced at 37°C and expressed in vivo2-4 days after infection, indicating that InvD is a virulence factor. During infection, InvD was expressed in all parts of the intestinal tract, but not in deeper lymphoid tissues. The crystal structure of the C-terminal adhesion domain of InvD revealed a distinct Ig-related fold, that, apart from the canonical β-sheets, comprises various modifications of and insertions into the Ig-core structure. We identified the Fab fragment of host-derived IgG/IgA antibodies as the target of the adhesion domain. Phage display panning and flow cytometry data further revealed that InvD exhibits a preferential binding specificity toward antibodies with VH3/VK1 variable domains and that it is specifically recruited to a subset of B cells. This finding suggests that InvD modulates Ig functions in the intestine and affects direct interactions with a subset of cell surface-exposed B-cell receptors. In summary, our results provide extensive insights into the structure of InvD and its specific interaction with the target molecule in the host.
ORIGINAL
Sadana et al.pdf
Sadana et al.pdf
after embargo allowed publisher's PDF
application/pdf
12519093
https://hzi.openrepository.com/bitstream/10033/621332/1/Sadana%20et%20al.pdf
a4028bbbb228fa8205fd6d95cf9b9ddd
MD5
1
true
CC-LICENSE
license_url
license_url
text/plain
49
https://hzi.openrepository.com/bitstream/10033/621332/2/license_url
924993ce0b3ba389f79f32a1b2735415
MD5
2
false
license_text
license_text
application/octet-stream
0
https://hzi.openrepository.com/bitstream/10033/621332/3/license_text
d41d8cd98f00b204e9800998ecf8427e
MD5
3
false
license_rdf
license_rdf
application/octet-stream
0
https://hzi.openrepository.com/bitstream/10033/621332/4/license_rdf
d41d8cd98f00b204e9800998ecf8427e
MD5
4
false
LICENSE
license.txt
license.txt
text/plain
1685
https://hzi.openrepository.com/bitstream/10033/621332/5/license.txt
cb598eeb10bfed09d26fd8d285172ad4
MD5
5
false
TEXT
Sadana et al.pdf.txt
Sadana et al.pdf.txt
Extracted Text
text/plain
117444
https://hzi.openrepository.com/bitstream/10033/621332/6/Sadana%20et%20al.pdf.txt
78dfa7e027ad033fccf7864f23dd9e93
MD5
6
false
THUMBNAIL
Sadana et al.pdf.jpg
Sadana et al.pdf.jpg
Generated Thumbnail
image/jpeg
113019
https://hzi.openrepository.com/bitstream/10033/621332/7/Sadana%20et%20al.pdf.jpg
5aa284dd8f4a2b40f421d255b6a9fd7a
MD5
7
false
10033/621332
oai:hzi.openrepository.com:10033/621332
2019-08-30 11:29:17.633
Helmholtz Zentrum für Infektionsforschung Repository
hzi@openrepository.com
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
oai:repository.helmholtz-hzi.de:10033/6219632019-10-01T07:08:45Zcom_10033_311308col_10033_559591col_10033_620561
Juettner, Norbert E.
dab75a4d01da24a23760d8919217524c
Schmelz, Stefan
91c14d4d519c8fc0f1446b2262626ea5
500
Anderl, Anita
15dd81a9b8c604c21cba055051d47409
500
Colin, Felix
a495b1b47335b0bb14bcc518239a9d5f
500
Classen, Moritz
cdb91c16e8f3b366d31b774eecb7102d
500
Pfeifer, Felicitas
d9c60d4d313b8ca23418cedfe1b9d133
500
Scrima, Andrea
c3ad854d78113b599fa24cb5e9e6fbcc
500
Fuchsbauer, Hans‐Lothar
b219fed9cdf8c0347355a786b823be12
HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.
2019-09-30T12:19:06Z
2019-09-30T12:19:06Z
2019-08-29
FEBS J. 2019 Aug 17. doi: 10.1111/febs.15044.
1742-464X
1742-4658
10.1111/febs.15044
http://hdl.handle.net/10033/621963
FEBS journal
Streptomyces mobaraensis is a key player for the industrial production of the protein cross-linking enzyme microbial transglutaminase (MTG). Extra-cellular activation of MTG by the transglutaminase-activating metalloprotease (TAMP) is regulated by the TAMP inhibitory protein SSTI that belongs to the large Streptomyces subtilisin inhibitor (SSI) family. Despite decades of SSI research, the binding site for metalloproteases such as TAMP remained elusive in most of the SSI proteins. Moreover, SSTI is a MTG substrate, and the preferred glutamine residues for SSTI cross-linking are not determined. To address both issues, that is, determination of the TAMP and the MTG glutamine binding sites, SSTI was modified by distinct point mutations as well as elongation or truncation of the N-terminal peptide by six and three residues respectively. Structural integrity of the mutants was verified by the determination of protein melting points and supported by unimpaired subtilisin inhibitory activity. While exchange of single amino acids could not disrupt decisively the SSTI TAMP interaction, the N-terminally shortened variants clearly indicated the highly conserved Leu40-Tyr41 as binding motif for TAMP. Moreover, enzymatic biotinylation revealed that an adjacent glutamine pair, upstream from Leu40-Tyr41 in the SSTI precursor protein, is the preferred binding site of MTG. This extension peptide disturbs the interaction with TAMP. The structure of SSTI was furthermore determined by X-ray crystallography. While no structural data could be obtained for the N-terminal peptide due to flexibility, the core structure starting from Tyr41 could be determined and analysed, which superposes well with SSI-family proteins. ENZYMES: Chymotrypsin, EC3.4.21.1; griselysin (SGMPII, SgmA), EC3.4.24.27; snapalysin (ScNP), EC3.4.24.77; streptogrisin-A (SGPA), EC3.4.21.80; streptogrisin-B (SGPB), EC3.4.21.81; subtilisin BPN', EC3.4.21.62; transglutaminase, EC2.3.2.13; transglutaminase-activating metalloprotease (TAMP), EC3.4.-.-; tri-/tetrapeptidyl aminopeptidase, EC3.4.11.-; trypsin, EC3.4.21.4. DATABASES: The atomic coordinates and structure factors (PDB 6I0I) have been deposited in the Protein Data Bank (http://www.rcsb.org).
en
Wiley
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
Cell Biology
Biochemistry
Molecular Biology
The N‐terminal peptide of the transglutaminase‐activating metalloprotease inhibitor from Streptomyces mobaraensis accommodates both inhibition and glutamine cross‐linking sites
Article
THUMBNAIL
2020-08-17
Juettner et al.pdf.jpg
Juettner et al.pdf.jpg
Generated Thumbnail
image/jpeg
45353
https://hzi.openrepository.com/bitstream/10033/621963/10/Juettner%20et%20al.pdf.jpg
9c4bd0395318d21e4ff18ed784623ca6
MD5
10
false
2020-08-17
Figure 1.tif.jpg
Figure 1.tif.jpg
IM Thumbnail
image/jpeg
30217
https://hzi.openrepository.com/bitstream/10033/621963/11/Figure%201.tif.jpg
56db01cc939f7ecddd55b9b4a0d7ad0a
MD5
11
false
2020-08-17
Figure 2.tif.jpg
Figure 2.tif.jpg
IM Thumbnail
image/jpeg
16045
https://hzi.openrepository.com/bitstream/10033/621963/12/Figure%202.tif.jpg
ec050768a6b725c2b06e3e7e0ffe1d21
MD5
12
false
2020-08-17
Figure 3.tiff.jpg
Figure 3.tiff.jpg
IM Thumbnail
image/jpeg
21100
https://hzi.openrepository.com/bitstream/10033/621963/13/Figure%203.tiff.jpg
ee06f1720fda7b62e30e4ba1c12fb17d
MD5
13
false
2020-08-17
Figure 4 .tif.jpg
Figure 4 .tif.jpg
IM Thumbnail
image/jpeg
28021
https://hzi.openrepository.com/bitstream/10033/621963/14/Figure%204%20.tif.jpg
1907d2c7b2ca4e4b8898ca0af6d4bcbb
MD5
14
false
figure5.tif.jpg
figure5.tif.jpg
IM Thumbnail
image/jpeg
60343
https://hzi.openrepository.com/bitstream/10033/621963/15/figure5.tif.jpg
d4cf8f1f0170fdf506d00df8043cfc7d
MD5
15
false
TEXT
2020-08-17
Juettner et al.pdf.txt
Juettner et al.pdf.txt
Extracted text
text/plain
51889
https://hzi.openrepository.com/bitstream/10033/621963/9/Juettner%20et%20al.pdf.txt
e5292a8237f1ea8be1853daafbaac6a9
MD5
9
false
LICENSE
license.txt
license.txt
text/plain
1685
https://hzi.openrepository.com/bitstream/10033/621963/8/license.txt
cb598eeb10bfed09d26fd8d285172ad4
MD5
8
false
CC-LICENSE
license_rdf
license_rdf
application/rdf+xml; charset=utf-8
1031
https://hzi.openrepository.com/bitstream/10033/621963/7/license_rdf
934f4ca17e109e0a05eaeaba504d7ce4
MD5
7
false
ORIGINAL
Juettner et al.pdf
Juettner et al.pdf
original manuscript
application/pdf
618081
https://hzi.openrepository.com/bitstream/10033/621963/1/Juettner%20et%20al.pdf
0dc9ef77cb65f32f31c32d30756afdba
MD5
1
true
Figure 1.tif
Figure 1.tif
figure 1
image/tiff
2206128
https://hzi.openrepository.com/bitstream/10033/621963/2/Figure%201.tif
b121f5a55a6baba701ce13beda766b07
MD5
2
false
Figure 2.tif
Figure 2.tif
figure 2
image/tiff
311286
https://hzi.openrepository.com/bitstream/10033/621963/3/Figure%202.tif
cc80274fde34601881c0e86c723ebbc2
MD5
3
false
Figure 3.tiff
Figure 3.tiff
figure 3
image/tiff
13885906
https://hzi.openrepository.com/bitstream/10033/621963/4/Figure%203.tiff
7a2f7952081015aea2f678c2ef7ea0ec
MD5
4
false
Figure 4 .tif
Figure 4 .tif
figure 4
image/tiff
837676
https://hzi.openrepository.com/bitstream/10033/621963/5/Figure%204%20.tif
4ef9ab2c8061d73cdab9bf27a771ff0b
MD5
5
false
figure5.tif
figure5.tif
figure 5
image/tiff
8904644
https://hzi.openrepository.com/bitstream/10033/621963/6/figure5.tif
b7884ce48f18fab757c13f47feb64496
MD5
6
false
10033/621963
oai:hzi.openrepository.com:10033/621963
2019-10-01 07:08:45.072
Helmholtz Zentrum für Infektionsforschung Repository
hzi@openrepository.com
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
oai:repository.helmholtz-hzi.de:10033/6226882021-01-29T11:12:46Zcom_10033_311308col_10033_559591col_10033_620561
Juettner, Norbert E
51015be037fa3943c3180a324d0d5251
Schmelz, Stefan
91c14d4d519c8fc0f1446b2262626ea5
500
Anderl, Anita
15dd81a9b8c604c21cba055051d47409
500
Colin, Felix
a495b1b47335b0bb14bcc518239a9d5f
500
Classen, Moritz
cdb91c16e8f3b366d31b774eecb7102d
500
Pfeifer, Felicitas
d9c60d4d313b8ca23418cedfe1b9d133
500
Scrima, Andrea
c3ad854d78113b599fa24cb5e9e6fbcc
500
Fuchsbauer, Hans-Lothar
be4a81b2a16b231e991ac87fffdc1f69
500
HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.
2021-01-19T11:41:53Z
2021-01-19T11:41:53Z
2019-08-29
FEBS J. 2020 Feb;287(4):708-720. doi: 10.1111/febs.15044. Epub 2019 Aug 29.
31420998
10.1111/febs.15044
http://hdl.handle.net/10033/622688
1742-4658
The FEBS journal
Streptomyces mobaraensis is a key player for the industrial production of the protein cross-linking enzyme microbial transglutaminase (MTG). Extra-cellular activation of MTG by the transglutaminase-activating metalloprotease (TAMP) is regulated by the TAMP inhibitory protein SSTI that belongs to the large Streptomyces subtilisin inhibitor (SSI) family. Despite decades of SSI research, the binding site for metalloproteases such as TAMP remained elusive in most of the SSI proteins. Moreover, SSTI is a MTG substrate, and the preferred glutamine residues for SSTI cross-linking are not determined. To address both issues, that is, determination of the TAMP and the MTG glutamine binding sites, SSTI was modified by distinct point mutations as well as elongation or truncation of the N-terminal peptide by six and three residues respectively. Structural integrity of the mutants was verified by the determination of protein melting points and supported by unimpaired subtilisin inhibitory activity. While exchange of single amino acids could not disrupt decisively the SSTI TAMP interaction, the N-terminally shortened variants clearly indicated the highly conserved Leu40-Tyr41 as binding motif for TAMP. Moreover, enzymatic biotinylation revealed that an adjacent glutamine pair, upstream from Leu40-Tyr41 in the SSTI precursor protein, is the preferred binding site of MTG. This extension peptide disturbs the interaction with TAMP. The structure of SSTI was furthermore determined by X-ray crystallography. While no structural data could be obtained for the N-terminal peptide due to flexibility, the core structure starting from Tyr41 could be determined and analysed, which superposes well with SSI-family proteins. ENZYMES: Chymotrypsin, EC3.4.21.1; griselysin (SGMPII, SgmA), EC3.4.24.27; snapalysin (ScNP), EC3.4.24.77; streptogrisin-A (SGPA), EC3.4.21.80; streptogrisin-B (SGPB), EC3.4.21.81; subtilisin BPN', EC3.4.21.62; transglutaminase, EC2.3.2.13; transglutaminase-activating metalloprotease (TAMP), EC3.4.-.-; tri-/tetrapeptidyl aminopeptidase, EC3.4.11.-; trypsin, EC3.4.21.4. DATABASES: The atomic coordinates and structure factors (PDB 6I0I) have been deposited in the Protein Data Bank (http://www.rcsb.org).
en
FEBS Press
Attribution 4.0 International
http://creativecommons.org/licenses/by-NC-ND/4.0
Streptomyces mobaraensis
crystal structure
metalloprotease inhibitor
serine protease inhibitor
transglutaminase
The N-terminal peptide of the transglutaminase-activating metalloprotease inhibitor from Streptomyces mobaraensis accommodates both inhibition and glutamine cross-linking sites.
Article
287
4
708
720
The FEBS journal
England
2021-01-19T11:41:55Z
THUMBNAIL
Juettner et al.pdf.jpg
Juettner et al.pdf.jpg
Generated Thumbnail
image/jpeg
103377
https://repository.helmholtz-hzi.de/bitstream/10033/622688/5/Juettner%20et%20al.pdf.jpg
0c4931bfe2d1cb777d24475166bf1111
MD5
5
false
TEXT
Juettner et al.pdf.txt
Juettner et al.pdf.txt
Extracted text
text/plain
53574
https://repository.helmholtz-hzi.de/bitstream/10033/622688/4/Juettner%20et%20al.pdf.txt
0e4c00977174efa0f33f3f8d256a2f24
MD5
4
false
LICENSE
license.txt
license.txt
text/plain
1685
https://repository.helmholtz-hzi.de/bitstream/10033/622688/3/license.txt
cb598eeb10bfed09d26fd8d285172ad4
MD5
3
false
CC-LICENSE
license_rdf
license_rdf
application/rdf+xml; charset=utf-8
908
https://repository.helmholtz-hzi.de/bitstream/10033/622688/2/license_rdf
0175ea4a2d4caec4bbcc37e300941108
MD5
2
false
ORIGINAL
Juettner et al.pdf
Juettner et al.pdf
delayed Open Access publication
application/pdf
2493709
https://repository.helmholtz-hzi.de/bitstream/10033/622688/1/Juettner%20et%20al.pdf
c2eb8334b82a2474d19c667e6464f99e
MD5
1
true
10033/622688
oai:repository.helmholtz-hzi.de:10033/622688
2021-01-29 11:12:46.778
Helmholtz Zentrum für Infektionsforschung Repository
hzi@openrepository.com
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