{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Zaric M"],"funding":["The Slovenian Research and Innovation Agency"],"pagination":["11910"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC12732864"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["26(24)"],"pubmed_abstract":["Cathepsin B (CatB) is a lysosomal cysteine protease that plays a major role in various pathologies and is therefore considered a valuable therapeutic target. To address species-specific inhibitor challenges, we characterized the selective binding of designed ankyrin repeat protein (DARPin) 4m3 toward mouse cathepsin B (mCatB) over human CatB (hCatB). The mCatB-DARPin 4m3 complex was validated by size-exclusion chromatography (SEC), nano-differential scanning fluorimetry (nano-DSF), and surface plasmon resonance (SPR), revealing high affinity binding (K<sub>D</sub> = 65.7 nM) and potent inhibition (Ki = 26.7 nM; mixed competitive/noncompetitive). DARPin 4m3 showed no binding/inhibition toward hCatB. The 1.67 Å crystal structure of the complex-the first for mCatB-identified key interaction r"],"journal":["International journal of molecular sciences"],"pubmed_title":["Structural and Proteomic Analysis of the Mouse Cathepsin B-DARPin 4m3 Complex Reveals Species-Specific Binding Determinants."],"pmcid":["PMC12732864"],"funding_grant_id":["P1-0140; P1-0048"],"pubmed_authors":["Novak M","Turk B","Gevaert K","Usenik A","Turk D","Kramer L","Vasiljeva O","Impens F","Tusar L","Zaric M"],"additional_accession":[]},"is_claimable":false,"name":"Structural and Proteomic Analysis of the Mouse Cathepsin B-DARPin 4m3 Complex Reveals Species-Specific Binding Determinants.","description":"Cathepsin B (CatB) is a lysosomal cysteine protease that plays a major role in various pathologies and is therefore considered a valuable therapeutic target. To address species-specific inhibitor challenges, we characterized the selective binding of designed ankyrin repeat protein (DARPin) 4m3 toward mouse cathepsin B (mCatB) over human CatB (hCatB). The mCatB-DARPin 4m3 complex was validated by size-exclusion chromatography (SEC), nano-differential scanning fluorimetry (nano-DSF), and surface plasmon resonance (SPR), revealing high affinity binding (K<sub>D</sub> = 65.7 nM) and potent inhibition (Ki = 26.7 nM; mixed competitive/noncompetitive). DARPin 4m3 showed no binding/inhibition toward hCatB. The 1.67 Å crystal structure of the complex-the first for mCatB-identified key interaction r","dates":{"release":"2025-01-01T00:00:00Z","publication":"2025 Dec","modification":"2026-05-28T03:21:15.918Z","creation":"2026-05-28T03:11:44.65Z"},"accession":"S-EPMC12732864","cross_references":{"pubmed":["41465336"],"doi":["10.3390/ijms262411910"]}}