Structure of the Dispase Autolysis-inducing Protein from Streptomyces mobaraensis and Glutamine Cross-linking Sites for Transglutaminase.

2.50
Hdl Handle:
http://hdl.handle.net/10033/621321
Title:
Structure of the Dispase Autolysis-inducing Protein from Streptomyces mobaraensis and Glutamine Cross-linking Sites for Transglutaminase.
Authors:
Fiebig, David; Schmelz, Stefan; Zindel, Stephan; Ehret, Vera; Beck, Jan; Ebenig, Aileen; Ehret, Marina; Fröls, Sabrina; Pfeifer, Felicitas; Kolmar, Harald; Fuchsbauer, Hans-Lothar; Scrima, Andrea
Abstract:
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.
Affiliation:
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Citation:
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.
Journal:
The Journal of biological chemistry
Issue Date:
2016
URI:
http://hdl.handle.net/10033/621321
DOI:
10.1074/jbc.M116.731109
PubMed ID:
27493205
PubMed Central ID:
PMC5034039
Additional Links:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5034039/
Type:
Article
Language:
en
ISSN:
1083-351X
Appears in Collections:
publications of the NG Structurbiologie der Autophagie (SBAU)

Full metadata record

DC FieldValue Language
dc.contributor.authorFiebig, Daviden
dc.contributor.authorSchmelz, Stefanen
dc.contributor.authorZindel, Stephanen
dc.contributor.authorEhret, Veraen
dc.contributor.authorBeck, Janen
dc.contributor.authorEbenig, Aileenen
dc.contributor.authorEhret, Marinaen
dc.contributor.authorFröls, Sabrinaen
dc.contributor.authorPfeifer, Felicitasen
dc.contributor.authorKolmar, Haralden
dc.contributor.authorFuchsbauer, Hans-Lotharen
dc.contributor.authorScrima, Andreaen
dc.date.accessioned2018-03-12T15:28:28Z-
dc.date.available2018-03-12T15:28:28Z-
dc.date.issued2016-
dc.identifier.citationStructure of the Dispase Autolysis-inducing Protein from Streptomyces mobaraensis and Glutamine Cross-linking Sites for Transglutaminase. 2016, 291 (39):20417-26 J. Biol. Chem.en
dc.identifier.issn1083-351X-
dc.identifier.pmid27493205-
dc.identifier.doi10.1074/jbc.M116.731109-
dc.identifier.urihttp://hdl.handle.net/10033/621321-
dc.description.abstractTransglutaminase 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
dc.language.isoenen
dc.relation.urlhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5034039/en
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.subject.meshBacterial Proteinsen
dc.subject.meshEscherichia colien
dc.subject.meshRecombinant Proteinsen
dc.subject.meshStreptomycesen
dc.subject.meshTransglutaminasesen
dc.titleStructure of the Dispase Autolysis-inducing Protein from Streptomyces mobaraensis and Glutamine Cross-linking Sites for Transglutaminase.en
dc.typeArticleen
dc.contributor.departmentHelmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.en
dc.identifier.journalThe Journal of biological chemistryen
dc.identifier.pmcidPMC5034039-

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