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Elastin degradation by cathepsin V requires two exosites.

The Journal of biological chemistry (2013-10-15)
Xin Du, Nelson L H Chen, Andre Wong, Charles S Craik, Dieter Brömme
ABSTRACT

Cathepsin V is a highly effective elastase and has been implicated in physiological and pathological extracellular matrix degradation. However, its mechanism of action remains elusive. Whereas human cathepsin V exhibits a potent elastolytic activity, the structurally homologous cathepsin L, which shares a 78% amino acid sequence, has only a minimal proteolytic activity toward insoluble elastin. This suggests that there are distinct structural domains that play an important role in elastinolysis. In this study, a total of 11 chimeras of cathepsins V and L were generated to identify elastin-binding domains in cathepsin V. Evaluation of these chimeras revealed two exosites contributing to the elastolytic activity of cathepsin V that are distant from the active cleft of the protease and are located in surface loop regions. Replacement of exosite 1 or 2 with analogous residues from cathepsin L led to a 75 and 43% loss in the elastolytic activity, respectively. Replacement of both exosites yielded a non-elastase variant similar to that of cathepsin L. Identification of these exosites may contribute to the design of inhibitors that will only affect the elastolytic activity of cysteine cathepsins without interfering with other physiological protease functions.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Elastin, soluble from bovine neck ligament, salt-free, lyophilized powder
Sigma-Aldrich
Elastase from porcine pancreas, Type III, lyophilized powder, Protein 55-85 %, ≥4.0 units/mg protein
Sigma-Aldrich
Elastase from porcine pancreas, lyophilized powder, suitable for cell culture
Sigma-Aldrich
Elastase from porcine pancreas, Type IV, Protein 50-90 %, lyophilized powder, ≥4.0 units/mg protein (biuret)
Sigma-Aldrich
Elastin from bovine neck ligament, powder
Sigma-Aldrich
Elastase from porcine pancreas, Type I, ≥4.0 units/mg protein
Sigma-Aldrich
Cathepsin L from human liver, ≥0.5 units/mg protein, solution