{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T05:02:47Z","timestamp":1791262967958,"version":"4.1.0"},"reference-count":0,"publisher":"Portland Press Ltd.","issue":"2","content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2003,10,15]]},"abstract":"<jats:p>The limited availability of highly selective cathepsin substrates seriously impairs studies designed to monitor individual cathepsin activities in biological samples. Among mammalian cysteine proteases, cathepsin K has a unique preference for a proline residue at P2, the primary determinant of its substrate specificity. Interestingly, congopain from Trypanosoma congolense also accommodates a proline residue in its S2 subsite. Analysis of a congopain model showed that amino acids forming its S2 subsite are identical with those of cathepsin K, except Leu67 which is replaced by a tyrosine residue in cathepsin K. Furthermore, amino acid residues of the congopain S2\u2032 binding pocket, which accepts a proline residue, are strictly identical with those of cathepsin K. Abz-HPGGPQ-EDN2ph [where Abz represents o-aminobenzoic acid and EDN2ph (=EDDnp) represents N-(2,4-dinitrophenyl)-ethylenediamine], a substrate initially developed for trypanosomal enzymes, was efficiently cleaved at the Gly\u2013Gly bond by cathepsin K (kcat\/Km=426000 M\u22121\u00b7s\u22121). On the other hand, Abz-HPGGPQ-EDN2ph was resistant to hydrolysis by cathepsins B, F, H, L, S and V (20 nM enzyme concentration) and the Y67L (Tyr67\u2192Leu)\/L205A cathepsin K mutant (20 nM), but still acted as a competitive inhibitor. Taken together, the selectivity of Abz-HPGGPQ-EDN2ph to cathepsin K primarily depends on the S2 and S2\u2032 subsite specificities of cathepsin K and the ionization state of histidine at P3. Whereas Abz-HPGGPQ-EDN2ph was hydrolysed by wild-type mouse fibroblast lysates, its hydrolysis was completely abolished in the cathepsin K-deficient samples, indicating that Abz-HPGGPQ-EDN2ph can be used to monitor selectively cathepsin K activity in physiological fluids and cell lysates.<\/jats:p>","DOI":"10.1042\/bj20030468","type":"journal-article","created":{"date-parts":[[2003,10,13]],"date-time":"2003-10-13T09:32:29Z","timestamp":1066037549000},"page":"307-312","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":53,"title":["Probing cathepsin K activity with a selective substrate spanning its active site"],"prefix":"10.1042","volume":"375","author":[{"given":"Fabien","family":"LECAILLE","sequence":"first","affiliation":[{"name":"INSERM EMI-U 00-10 \u2018Prot\u00e9ases et Vectorisation\u2019, Laboratoire d'Enzymologie et Chimie des Prot\u00e9ines, Facult\u00e9 de M\u00e9decine, Universit\u00e9 Fran\u00e7ois Rabelais, 2 bis, Boulevard Tonnell\u00e9, F-37032 Tours Cedex, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Enrico","family":"WEIDAUER","sequence":"additional","affiliation":[{"name":"Department of Human Genetics, Mount Sinai School of Medicine, New York, NY 10029, U.S.A."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria A.","family":"JULIANO","sequence":"additional","affiliation":[{"name":"Escola Paulista de Medicina, Universidade Federal de S\u00e3o Paulo, 04044-020 S\u00e3o Paulo, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dieter","family":"BR\u00d6MME","sequence":"additional","affiliation":[{"name":"Department of Human Genetics, Mount Sinai School of Medicine, New York, NY 10029, U.S.A."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gilles","family":"LALMANACH","sequence":"additional","affiliation":[{"name":"INSERM EMI-U 00-10 \u2018Prot\u00e9ases et Vectorisation\u2019, Laboratoire d'Enzymologie et Chimie des Prot\u00e9ines, Facult\u00e9 de M\u00e9decine, Universit\u00e9 Fran\u00e7ois Rabelais, 2 bis, Boulevard Tonnell\u00e9, F-37032 Tours Cedex, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"288","container-title":["Biochemical Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/375\/2\/307\/712098\/bj3750307.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/375\/2\/307\/712098\/bj3750307.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,11,22]],"date-time":"2021-11-22T12:38:03Z","timestamp":1637584683000},"score":1,"resource":{"primary":{"URL":"https:\/\/portlandpress.com\/biochemj\/article\/375\/2\/307\/40750\/Probing-cathepsin-K-activity-with-a-selective"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2003,10,15]]},"references-count":0,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2003,10,15]]}},"URL":"https:\/\/doi.org\/10.1042\/bj20030468","relation":{},"ISSN":["0264-6021","1470-8728"],"issn-type":[{"value":"0264-6021","type":"print"},{"value":"1470-8728","type":"electronic"}],"subject":[],"published":{"date-parts":[[2003,10,15]]}}}