6JD0
Structure of mutant human cathepsin L, engineered for GAG binding
Summary for 6JD0
Entry DOI | 10.2210/pdb6jd0/pdb |
Descriptor | Cathepsin L1, GLYCEROL, CHLORIDE ION, ... (10 entities in total) |
Functional Keywords | cathepsin l, collagenase, gag, hydrolase |
Biological source | Homo sapiens (Human) |
Total number of polymer chains | 1 |
Total formula weight | 45008.26 |
Authors | Choudhury, D.,Biswas, S. (deposition date: 2019-01-30, release date: 2020-02-05, Last modification date: 2024-10-30) |
Primary citation | Choudhury, D.,Biswas, S. Structure-guided protein engineering of human cathepsin L for efficient collagenolytic activity. Protein Eng.Des.Sel., 34:-, 2021 Cited by PubMed Abstract: Engineering precise substrate specificity of proteases advances the potential to use them in biotechnological and therapeutic applications. Collagen degradation, a physiological process mediated by collagenases, is an integral part of extracellular matrix remodeling and when uncontrolled, implicated in different pathological conditions. Lysosomal cathepsin-K cleaves triple helical collagen fiber, whereas cathepsin-L cannot do so. In this study, we have imparted collagenolytic property to cathepsin-L, by systematically engineering proline-specificity and glycosaminoglycans (GAG)-binding surface in the protease. The proline-specific mutant shows high specificity for prolyl-peptidic substrate but is incapable of cleaving collagen. Engineering a GAG-binding surface on the proline-specific mutant enabled it to degrade type-I collagen in the presence of chondroitin-4-sulfate (C4-S). We also present the crystal structures of proline-specific (1.4 Å) and collagen-specific (1.8 Å) mutants. Finally docking studies with prolyl-peptidic substrate (Ala-Gly-Pro-Arg-Ala) at the active site and a C4-S molecule at the GAG-binding site enable us to identify key structural features responsible for collagenolytic activity of cysteine cathepsins. PubMed: 33825882DOI: 10.1093/protein/gzab005 PDB entries with the same primary citation |
Experimental method | X-RAY DIFFRACTION (1.805 Å) |
Structure validation
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