{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,9]],"date-time":"2026-01-09T03:11:19Z","timestamp":1767928279758,"version":"3.49.0"},"reference-count":53,"publisher":"Wiley","issue":"13","license":[{"start":{"date-parts":[[2016,6,6]],"date-time":"2016-06-06T00:00:00Z","timestamp":1465171200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001824","name":"Grantov\u00e1 Agentura \u010cesk\u00e9 Republiky","doi-asserted-by":"publisher","award":["P208\u201012\u2010G016"],"award-info":[{"award-number":["P208\u201012\u2010G016"]}],"id":[{"id":"10.13039\/501100001824","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["febs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The FEBS Journal"],"published-print":{"date-parts":[[2016,7]]},"abstract":"Glutamate carboxypeptidase III<\/jats:styled-content> (GCPIII<\/jats:styled-content>) is best known as a homologue of glutamate carboxypeptidase II<\/jats:styled-content> [GCPII<\/jats:styled-content>; also known as prostate\u2010specific membrane antigen (PSMA<\/jats:styled-content>)], a protease involved in neurological disorders and overexpressed in a number of solid cancers. However, mouse GCPIII<\/jats:styled-content> was recently shown to cleave \u03b2\u2010citrylglutamate (BCG<\/jats:styled-content>), suggesting that these two closely related enzymes have distinct functions. To develop a tool to dissect, evaluate and quantify the activities of human GCPII<\/jats:styled-content> and GCPIII<\/jats:styled-content>, we analysed the catalytic efficiencies of these enzymes towards three physiological substrates. We observed a high efficiency of BCG<\/jats:styled-content> cleavage by GCPIII<\/jats:styled-content> but not GCPII<\/jats:styled-content>. We also identified a strong modulation of GCPIII<\/jats:styled-content> enzymatic activity by divalent cations, while we did not observe this effect for GCPII<\/jats:styled-content>. Additionally, we used X\u2010ray crystallography and computational modelling (quantum and molecular mechanical calculations) to describe the mechanism of BCG<\/jats:styled-content> binding to the active sites of GCPII<\/jats:styled-content> and GCPIII<\/jats:styled-content>, respectively. Finally, we took advantage of the substantial differences in the enzymatic efficiencies of GCPII<\/jats:styled-content> and GCPIII<\/jats:styled-content> towards their substrates, using enzymatic assays for specific detection of these proteins in human tissues. Our findings suggest that GCPIII<\/jats:styled-content> may not act merely as a complementary enzyme to GCPII<\/jats:styled-content>, and it more likely possesses a specific physiological function related to BCG<\/jats:styled-content> metabolism in the human body.<\/jats:p><\/jats:sec>Database<\/jats:title>The X\u2010ray structure of GCPII<\/jats:styled-content> Glu424Ala in complex with BCG<\/jats:styled-content> has been deposited in the RCSB<\/jats:styled-content> Protein Data Bank under accession code 5F09<\/jats:ext-link>.<\/jats:p><\/jats:sec>","DOI":"10.1111\/febs.13761","type":"journal-article","created":{"date-parts":[[2016,5,24]],"date-time":"2016-05-24T01:54:11Z","timestamp":1464054851000},"page":"2528-2545","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":24,"title":["Comparison of human glutamate carboxypeptidases II<\/scp> and III<\/scp> reveals their divergent substrate 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