Crystal structure of TBC1D15 GTPase‐activating protein (GAP) domain and its activity on Rab GTPases
TBC1D15 belongs to the TBC (Tre‐2/Bub2/Cdc16) domain family and functions as a GTPase‐activating protein (GAP) for Rab GTPases. So far, the structure of TBC1D15 or the TBC1D15·Rab complex has not been determined, thus, its catalytic mechanism on Rab GTPases is still unclear. In this study, we solved...
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Published in | Protein science Vol. 26; no. 4; pp. 834 - 846 |
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Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
Wiley Subscription Services, Inc
01.04.2017
John Wiley and Sons Inc |
Subjects | |
Online Access | Get full text |
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Summary: | TBC1D15 belongs to the TBC (Tre‐2/Bub2/Cdc16) domain family and functions as a GTPase‐activating protein (GAP) for Rab GTPases. So far, the structure of TBC1D15 or the TBC1D15·Rab complex has not been determined, thus, its catalytic mechanism on Rab GTPases is still unclear. In this study, we solved the crystal structures of the Shark and Sus TBC1D15 GAP domains, to 2.8 Å and 2.5 Å resolution, respectively. Shark‐TBC1D15 and Sus‐TBC1D15 belong to the same subfamily of TBC domain‐containing proteins, and their GAP‐domain structures are highly similar. This demonstrates the evolutionary conservation of the TBC1D15 protein family. Meanwhile, the newly determined crystal structures display new variations compared to the structures of yeast Gyp1p Rab GAP domain and TBC1D1. GAP assays show that Shark and Sus GAPs both have higher catalytic activity on Rab11a·GTP than Rab7a·GTP, which differs from the previous study. We also demonstrated the importance of arginine and glutamine on the catalytic sites of Shark GAP and Sus GAP. When arginine and glutamine are changed to alanine or lysine, the activities of Shark GAP and Sus GAP are lost. |
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Bibliography: | The atomic coordinates and structure factors of the Shark GAP and Sus GAP have been deposited in the PDB Accession number (PDB ID codes 5TUB and 5TUC). http://www.rcsb.org ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Accession number: The atomic coordinates and structure factors of the Shark GAP and Sus GAP have been deposited in the PDB (http://www.rcsb.org) (PDB ID codes 5TUB and 5TUC). |
ISSN: | 0961-8368 1469-896X |
DOI: | 10.1002/pro.3132 |