Crystal Structure and Conformational Dynamics of Pyrococcus furiosus Prolyl Oligopeptidase
Enzymes in the prolyl oligopeptidase family possess unique structures and substrate specificities that are important for their biological activity and for potential biocatalytic applications. The crystal structures of Pyrococcus furiosus (Pfu) prolyl oligopeptidase (POP) and the corresponding S477C...
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Published in | Biochemistry (Easton) Vol. 58; no. 12; pp. 1616 - 1626 |
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Main Authors | , , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
American Chemical Society
26.03.2019
American Chemical Society (ACS) |
Subjects | |
Online Access | Get full text |
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Summary: | Enzymes in the prolyl oligopeptidase family possess unique structures and substrate specificities that are important for their biological activity and for potential biocatalytic applications. The crystal structures of Pyrococcus furiosus (Pfu) prolyl oligopeptidase (POP) and the corresponding S477C mutant were determined to 1.9 and 2.2 Å resolution, respectively. The wild type enzyme crystallized in an open conformation, indicating that this state is readily accessible, and it contained bound chloride ions and a prolylproline ligand. These structures were used as starting points for molecular dynamics simulations of Pfu POP conformational dynamics. The simulations showed that large-scale domain opening and closing occurred spontaneously, providing facile substrate access to the active site. Movement of the loop containing the catalytically essential histidine into a conformation similar to those found in structures with fully formed catalytic triads also occurred. This movement was modulated by chloride binding, providing a rationale for experimentally observed activation of POP peptidase catalysis by chloride. Thus, the structures and simulations reported in this study, combined with existing biochemical data, provide a number of insights into POP catalysis. |
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Bibliography: | USDOE Office of Science (SC) Lilly Endowment, Inc. National Institutes of Health (NIH) US Army Research Laboratory (USARL) US Army Research Office (ARO) Indiana METACyt Initiative National Science Foundation (NSF) P41 GM103403; S10 RR029205; AC02-06CH11357; W911NF-14-1-0334; 66796-LS-RIP; CHE-1351991; W911NF-18-1-0200 The David and Lucile Packard Foundation National Institute of General Medical Sciences (NIGMS) Present Addresses R.B.: National Institute on Alcohol Abuse and Alcoholism (NIAAA), 5625 Fishers Ln room 3N15, Rockville, MD 20852 K.E.G.: University of California - Los Angeles, Boyer Hall 610, 611 Charles E. Young Drive East, Los Angeles, CA 90095 These authors contributed equally. |
ISSN: | 0006-2960 1520-4995 |
DOI: | 10.1021/acs.biochem.9b00031 |