{"doi":"10.1110/ps.062666607","title":"The S2 subsites of cathepsins K and L and their contribution to collagen degradation","abstract":"<jats:title>Abstract</jats:title><jats:p>The exchange of residues 67 and 205 of the S2 pocket of human cysteine cathepsins K and L induces a permutation of their substrate specificity toward fluorogenic peptide substrates. While the cathepsin L‐like cathepsin K (Tyr67Leu/Leu205Ala) mutant has a marked preference for Phe, the Leu67Tyr/Ala205Leu cathepsin L variant shows an effective cathepsin K‐like preference for Leu and Pro. A similar turnaround of inhibition was observed by using specific inhibitors of cathepsin K [1‐(<jats:italic>N</jats:italic>‐Benzyloxycarbonyl‐leucyl)‐5‐(<jats:italic>N</jats:italic>‐Boc‐phenylalanyl‐leucyl)carbohydrazide] and cathepsin L [<jats:italic>N</jats:italic>‐(4‐biphenylacetyl)‐<jats:italic>S</jats:italic>‐methylcysteine‐(D)‐Arg‐Phe‐β‐phenethylamide]. Molecular modeling studies indicated that mutations alter the character of both S2 and S3 subsites, while docking calculations were consistent with kinetics data. The cathepsin K‐like cathepsin L was unable to mimic the collagen‐degrading activity of cathepsin K against collagens I and II, DQ‐collagens I and IV, and elastin‐Congo Red. In summary, double mutations of the S2 pocket of cathepsins K (Y67L/L205A) and L (L67Y/A205L) induce a switch of their enzymatic specificity toward small selective inhibitors and peptidyl substrates, confirming the key role of residues 67 and 205. However, mutations in the S2 subsite pocket of cathepsin L alone without engineering of binding sites to chondroitin sulfate are not sufficient to generate a cathepsin K‐like collagenase, emphasizing the pivotal role of the complex formation between glycosaminoglycans and cathepsin K for its unique collagenolytic activity.</jats:p>","journal":"Protein Science","year":2007,"id":34760,"datarank":2.330047561750596,"base_score":4.04305126783455,"endowment":4.04305126783455,"self_citation_contribution":0.6064576901751826,"citation_network_contribution":1.7235898715754132,"self_endowment_contribution":0.6064576901751826,"citer_contribution":1.7235898715754132,"corpus_percentile":null,"corpus_rank":null,"citation_count":56,"citer_count":41,"citers_with_citation_signal":41,"citers_with_endowment":41,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":179469,"name":"Shafinaz Chowdhury","orcid":null,"position":1,"is_corresponding":false},{"id":174539,"name":"Enrico Purisima","orcid":null,"position":2,"is_corresponding":false},{"id":90287,"name":"Dieter Brömme","orcid":"0000-0003-0517-5224","position":3,"is_corresponding":false},{"id":179471,"name":"Gilles Lalmanach","orcid":null,"position":4,"is_corresponding":false},{"id":179467,"name":"Fabien Lecaille","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"base_score":4.04305126783455,"endowment":4.04305126783455,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"17384231","pmcid":"PMC2203344","openalex_id":"https://openalex.org/W2063716200","authors":[],"funders":[],"total_grants":0,"fwci":1.9928,"citation_percentile":0.8827832,"influential_citations":3,"citation_trend":[{"year":2012,"count":3},{"year":2013,"count":2},{"year":2014,"count":1},{"year":2015,"count":4},{"year":2016,"count":2},{"year":2017,"count":4},{"year":2018,"count":2},{"year":2019,"count":3},{"year":2020,"count":4},{"year":2021,"count":3},{"year":2022,"count":2},{"year":2023,"count":2},{"year":2024,"count":2},{"year":2026,"count":1}],"oa_status":"closed","license":"http://onlinelibrary.wiley.com/termsAndConditions#vor","oa_locations":[{"url":"https://europepmc.org/articles/pmc2203344?pdf=render","host_type":"GREEN"},{"url":"https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1110%2Fps.062666607","host_type":"publisher"},{"url":"https://onlinelibrary.wiley.com/doi/pdf/10.1110/ps.062666607","host_type":"publisher"},{"url":"https://doi.org/10.1110/ps.062666607","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/17384231","host_type":"repository"},{"url":"https://nrc-publications.canada.ca/fra/voir/objet/?id=41c2c289-701d-4992-9bb0-6243fdcfac22","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/2203344","host_type":"repository"}],"fields_of_study":["Protease and Inhibitor Mechanisms","Bone Metabolism and Diseases","Bone and Dental Protein Studies","Chemistry","Medicine","Biology"],"mesh_terms":["Cathepsins","Collagen","Cysteine Endopeptidases","Electrophoresis, Polyacrylamide Gel","Hydrolysis","Substrate Specificity","Cysteine Proteinase Inhibitors","Cathepsin K","Cathepsin L"],"keywords":["Cathepsin O","Cathepsin A","Cathepsin C","Cathepsin","Cathepsin E","Chemistry","Cathepsin H","Biochemistry","Cathepsin S","Cathepsin L","Cathepsin K","Cathepsin L1","Cathepsin B","Mutant","Molecular biology","Enzyme","Biology","Gene"],"sdg_mappings":[{"sdg_number":0,"sdg_label":"Clean water and sanitation"}],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[{"name":"pdb"}],"source":"live","citation_network_status":"fetched"},"created_at":"2026-06-09T20:02:35.511271Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}