Cleavage of type I procollagen by human mast cell chymase initiates collagen fibril formation and generates a unique carboxyl-terminal propeptide.
The ability of human mast cell chymase and tryptase to process procollagen was examined. Purified human intestinal smooth muscle cell procollagen was incubated with human mast cell tryptase or human mast cell chymase. Purified chymase, but not tryptase, exhibited procollagen proteinase activity in the presence of EDTA. Addition of purified porcine heparin over a range of 0.1-100 microg/ml did not affect either the rate or the products of procollagen chymase cleavage. The cleavage site of chymase on the pro-alpha1(I) collagen carboxyl terminus was found to be in the propeptide region at Leu-1248-Ser-1249. Cleavage at this site suggested that the collagen products would form fibrils and confirmed the production of a unique carboxyl-terminal propeptide. Turbidometric fibril formation assay demonstrated de novo formation of chymase-generated collagen fibrils with characteristic lag, growth, and plateau phases. When observed by dark field microscopy, these fibrils were similar to fibrils formed by the action of procollagen proteinases. Thus, mast cell chymase, but not tryptase, exhibits procollagen peptidase-like activity as evidenced by its ability to process procollagen to fibril-forming collagen with concurrent formation of a unique carboxyl-terminal propeptide. These data demonstrate that mast cell chymase has a potential role in the regulation of collagen biosynthesis and in the pathogenesis of fibrosis.
Kofford
M W
MW
Department of Pediatrics, Medical College of Virginia/Virginia Commonwealth University, Richmond, Virginia 23298-0529, USA. mkofford@GEMS.vcu.edu
Schwartz
L B
LB
Schechter
N M
NM
Yager
D R
DR
Diegelmann
R F
RF
Graham
M F
MF
D000595:Amino Acid Sequence; D053818:Chymases; D003094:Collagen / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D008407:Mast Cells / Q000378:metabolism*; D008969:Molecular Sequence Data; D011347:Procollagen / Q000378:metabolism* / Q000648:ultrastructure; D012697:Serine Endopeptidases / Q000378:metabolism*
D011347:Procollagen; D003094:Collagen; D012697:Serine Endopeptidases; D053818:Chymases