6JD8

Structure of a proline specific mutant of human cathepsin L

Summary for 6JD8

• Entry DOI: 10.2210/pdb6jd8/pdb
• Descriptor: Cathepsin L1, GLYCEROL, ETHANOL, ... (7 entities in total)
• Functional Keywords: protease, proline-specific, engineered, hydrolase
• Biological source: Homo sapiens (Human)
• Total number of polymer chains: 1
• Total formula weight: 41881.71

Authors

Choudhury, D.,Biswas, S. (deposition date: 2019-01-31, release date: 2020-02-05, Last modification date: 2023-11-29)

Primary citation

Choudhury, D.,Biswas, S.
Structure-guided protein engineering of human cathepsin L for efficient collagenolytic activity.
Protein Eng.Des.Sel., 34:-, 2021

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: 33825882
DOI: 10.1093/protein/gzab005

Experimental method

X-RAY DIFFRACTION (1.457 Å)