{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T15:14:00Z","timestamp":1772118840103,"version":"3.50.1"},"reference-count":40,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2025,9,11]],"date-time":"2025-09-11T00:00:00Z","timestamp":1757548800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100006769","name":"Russian Science Foundation","doi-asserted-by":"publisher","award":["22\u201025\u201000648"],"award-info":[{"award-number":["22\u201025\u201000648"]}],"id":[{"id":"10.13039\/501100006769","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Proteins"],"published-print":{"date-parts":[[2026,2]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Cysteine cathepsins have been suggested as attractive therapeutic targets due to their critical role in several pathologies. In particular, inhibitors of cysteine cathepsins reduce the viability of tumor cells. The present study uses enzyme kinetics to characterize the interaction of human cathepsins L and S with their peptide substrate acetyl\u2010QLLR\u20107\u2010amino\u20104\u2010methylcoumarin (Ac\u2010QLLR\u2010AMC) and peptide inhibitors with anti\u2010tumor activity: FFSFGGAL (CS\u2010PEP1) and acetyl\u2010PLVE\u2010fluoromethyl\u2010ketone (Ac\u2010PLVE\u2010fmk). Due to multiple cellular locations of cathepsins, our study is conducted under different pH conditions, simulating lysosomal and cytosolic environments (pH\u20094.6 and 6.5\u20137.0). Catalytic activities of both cathepsins are higher at pH 6.5\u20137.0 compared to pH\u20094.6. Affinities for the substrate or inhibitor CS\u2010PEP1 are higher for cathepsin L than S independent of pH, but show different pH sensitivities, reciprocating different pI's of the cathepsins. Mixed inhibition by CS\u2010PEP1 is demonstrated for both cathepsins. While preincubation of cathepsins with CS\u2010PEP1 does not enhance the inhibition, Ac\u2010PLVE\u2010fmk inactivates both cathepsins in the preincubation medium. A strong increase in the inactivation rate is observed with increasing pH in the interval including p\n                    <jats:italic>K<\/jats:italic>\n                    <jats:sub>a<\/jats:sub>\n                    of the active site cysteine residues of cathepsins, in agreement with the irreversible modification by mono\u2010fluoromethyl ketones of the catalytic thiolate anion. At pH\u20094.6, cathepsin L has a higher affinity for Ac\u2010PLVE\u2010fmk, but a slower rate of the irreversible modification compared to cathepsin S. Our findings highlight opportunities for differential targeting of L and S cathepsins by peptide inhibitors in different cellular compartments, providing directions for cathepsin\u2010 and location\u2010specific drug design.\n                  <\/jats:p>","DOI":"10.1002\/prot.70047","type":"journal-article","created":{"date-parts":[[2025,9,12]],"date-time":"2025-09-12T03:00:37Z","timestamp":1757646037000},"page":"582-597","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Kinetic Characterization of Inhibition of Cathepsins L and S by Peptides With Anticancer Potential"],"prefix":"10.1002","volume":"94","author":[{"given":"Olga E.","family":"Chepikova","sequence":"first","affiliation":[{"name":"Institute of Translational Medicine and Biotechnology Sechenov First Moscow State Medical University  Moscow Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Victoria I.","family":"Bunik","sequence":"additional","affiliation":[{"name":"Belozersky Institute of Physicochemical Biology Lomonosov Moscow State University  Moscow Russia"},{"name":"Faculty of Bioengineering and Bioinformatics Lomonosov Moscow State University  Moscow Russia"},{"name":"Department of Biochemistry Sechenov First Moscow State Medical University  Moscow Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ivan V.","family":"Rodionov","sequence":"additional","affiliation":[{"name":"Department of Biochemistry Sechenov First Moscow State Medical University  Moscow Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Neonila V.","family":"Gorokhovets","sequence":"additional","affiliation":[{"name":"Institute of Translational Medicine and Biotechnology Sechenov First Moscow State Medical University  Moscow 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