3H7D

The crystal structure of the cathepsin K Variant M5 in complex with chondroitin-4-sulfate

Summary for 3H7D

• Entry DOI: 10.2210/pdb3h7d/pdb
• Related: 3C9E
• Descriptor: Cathepsin K, 2-acetamido-2-deoxy-4-O-sulfo-beta-D-galactopyranose-(1-4)-beta-D-glucopyranuronic acid-(1-3)-2-acetamido-2-deoxy-4-O-sulfo-beta-D-galactopyranose-(1-4)-beta-D-glucopyranuronic acid-(1-3)-2-acetamido-2-deoxy-4-O-sulfo-beta-D-galactopyranose-(1-4)-beta-D-glucopyranuronic acid, CALCIUM ION, ... (5 entities in total)
• Functional Keywords: glycosaminoglycan, sulfhydryl peptidase, cathepsin k mutant, ternary complex, disease mutation, disulfide bond, glycoprotein, hydrolase, lysosome, protease, thiol protease, zymogen
• Biological source: Homo sapiens (human)
• Total number of polymer chains: 2
• Total formula weight: 49115.78

Authors

Cherney, M.M.,Kienetz, M.,Bromme, D.,James, M.N.G. (deposition date: 2009-04-24, release date: 2010-04-28, Last modification date: 2024-11-06)

Primary citation

Cherney, M.M.,Lecaille, F.,Kienitz, M.,Nallaseth, F.S.,Li, Z.,James, M.N.,Bromme, D.
Structure-activity analysis of cathepsin K/chondroitin 4-sulfate interactions.
J.Biol.Chem., 286:8988-8998, 2011

PubMed Abstract: In the presence of oligomeric chondroitin 4-sulfate (C4-S), cathepsin K (catK) forms a specific complex that was shown to be the source of the major collagenolytic activity in bone osteoclasts. C4-S forms multiple contacts with amino acid residues on the backside of the catK molecule that help to facilitate complex formation. As cathepsin L does not exhibit a significant collagenase activity in the presence or in the absence of C4-S, we substituted the C4-S interacting residues in catK with those of cathepsin L. Variants revealed altered collagenolytic activities with the largest inhibitory effect shown by the hexavariant M5. None of the variants showed a reduction in their gelatinolytic and peptidolytic activities when compared with wild-type catK, indicating no structural alteration within their active sites. However, the crystal structure of the M5 variant in the presence of oligomeric C4-S revealed a different binding of chondroitin 4-sulfate. C4-S is not continuously ordered as it is in the wild-type catK·C4-S complex. The orientation and the direction of the hexasaccharide on the catK surface have changed, so that the hexasaccharide is positioned between two symmetry-related molecules. Only one M5 variant molecule of the dimer that is present in the asymmetric unit interacts with C4-S. These substitutions have changed the mode of catK binding to C4-S and, as a result, have likely affected the collagenolytic potential of the variant. The data presented here support our hypothesis that distinct catK/C4-S interactions are necessary for the collagenolytic activity of the enzyme.

PubMed: 21193413
DOI: 10.1074/jbc.M110.126706

Experimental method

X-RAY DIFFRACTION (2.242 Å)