McLuskey, K., Grewal, J. S., Das, D., Godzik, A., Lesley, S. A., Deacon, A. M., Coombs, G. H., Elsliger, M.-A., Wilson, I. A. and Mottram, J. C. (2016) Crystal structure and activity studies of the C11 cysteine peptidase from Parabacteroides merdae in the human gut microbiome. Journal of Biological Chemistry, 291, pp. 9482-9491. (doi: 10.1074/jbc.M115.706143) (PMID:26940874) (PMCID:PMC4850288)
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Abstract
Clan CD cysteine peptidases, a structurally related group of peptidases that include mammalian caspases, exhibit a wide range of important functions, along with a variety of specificities and activation mechanisms. However, for the clostripain family (denoted C11), little is currently known. Here, we describe the first crystal structure of a C11 protein from the human gut bacterium, Parabacteroides merdae (PmC11), determined to 1.7 Å resolution. PmC11 is a monomeric cysteine peptidase that comprises an extended caspase-like α/β/α sandwich and an unusual C-terminal domain. It shares core structural elements with clan CD cysteine peptidases but otherwise structurally differs from the other families in the clan. These studies also revealed a well-ordered break in the polypeptide chain at K147, resulting in a large conformational rearrangement close to the active site. Biochemical and kinetic analysis revealed K147 to be an intramolecular processing site at which cleavage is required for full activation of the enzyme, suggesting an auto-inhibitory mechanism for self-preservation. PmC11 has an acidic binding pocket and a preference for basic substrates, and accepts both Arg and Lys substrates and does not require Ca2+ for activity. Collectively, these data provide insights into the mechanism and activity of PmC11 and a detailed framework for studies on C11 peptidases from other phylogenetic kingdoms.
| Item Type: | Articles |
|---|---|
| Status: | Published |
| Refereed: | Yes |
| Glasgow Author(s) Enlighten ID: | Coombs, Professor Graham and McLuskey, Dr Karen and Grewal, Dr Jaspreet Singh and Mottram, Professor Jeremy |
| Authors: | McLuskey, K., Grewal, J. S., Das, D., Godzik, A., Lesley, S. A., Deacon, A. M., Coombs, G. H., Elsliger, M.-A., Wilson, I. A., and Mottram, J. C. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Infection & Immunity |
| Journal Name: | Journal of Biological Chemistry |
| Publisher: | American Society for Biochemistry and Molecular Biology, Inc. |
| ISSN: | 0021-9258 |
| ISSN (Online): | 1083-351X |
| Published Online: | 03 March 2016 |
| Copyright Holders: | Copyright © 2016 The Authors |
| First Published: | First published in Journal of Biological Chemistry 291:9482-9491 |
| Publisher Policy: | Reproduced under a Creative Commons License |
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