Identification, biochemical characterization and crystallization of the central region of human ATG16L1.

2.50
Hdl Handle:
http://hdl.handle.net/10033/621157
Title:
Identification, biochemical characterization and crystallization of the central region of human ATG16L1.
Authors:
Archna, Archna; Scrima, Andrea
Abstract:
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.
Affiliation:
Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr.7, 38124 Braunschweig, Germany.
Citation:
Identification, biochemical characterization and crystallization of the central region of human ATG16L1. 2017, 73 (Pt 10):560-567 Acta Crystallogr F Struct Biol Commun
Journal:
Acta crystallographica. Section F, Structural biology communications
Issue Date:
1-Oct-2017
URI:
http://hdl.handle.net/10033/621157
DOI:
10.1107/S2053230X17013280
PubMed ID:
28994404
Type:
Article
Language:
en
ISSN:
2053-230X
Appears in Collections:
research group of structural biology of autophagy (SBAU)

Full metadata record

DC FieldValue Language
dc.contributor.authorArchna, Archnaen
dc.contributor.authorScrima, Andreaen
dc.date.accessioned2017-11-03T10:13:04Z-
dc.date.available2017-11-03T10:13:04Z-
dc.date.issued2017-10-01-
dc.identifier.citationIdentification, biochemical characterization and crystallization of the central region of human ATG16L1. 2017, 73 (Pt 10):560-567 Acta Crystallogr F Struct Biol Communen
dc.identifier.issn2053-230X-
dc.identifier.pmid28994404-
dc.identifier.doi10.1107/S2053230X17013280-
dc.identifier.urihttp://hdl.handle.net/10033/621157-
dc.description.abstractATG16L1 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
dc.language.isoenen
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.titleIdentification, biochemical characterization and crystallization of the central region of human ATG16L1.en
dc.typeArticleen
dc.contributor.departmentHelmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr.7, 38124 Braunschweig, Germany.en
dc.identifier.journalActa crystallographica. Section F, Structural biology communicationsen

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