Susceptibility of type I collagen containing mutated alpha 1(1) chains to cleavage by human neutrophil collagenase.
Two members of the matrix metalloproteinase family which can cleave native types I, II and III triple helical collagens are collagenases from fibroblasts and neutrophils. These enzymes are the products of different genes which share structural motifs but are only 57% identical. In this study, we determined the site of cleavage in the alpha 1(I) chains and showed that the neutrophil collagenase acted at the same site as the fibroblast collagenase. We also used collagens as substrates which were generated by site-directed mutagenesis of the murine Col1a1 gene and found that the pattern of susceptibility to cleavage by purified neutrophil collagenase was indistinguishable from that previously described for the fibroblast collagenase. Collagens containing substitutions of Pro for Ile-776 (P1) were not cleaved; whereas those containing substitutions of Met for Ile-776 were cleaved. Type I collagen which contained alpha 1(I) chains in which there were double substitutions of Pro for Gln-774 (P2) and Ala-777 (P2') were also not cleaved. These type I collagens contained wild type alpha 2(I) chains as well as mutant alpha 1(I) chains in the mixed helical trimers; the alpha 2(I) chain in the trimers containing the resistant alpha 1(I) chains were also not cleaved by the neutrophil collagenase.
Hasty
K A
KA
Department of Anatomy and Neurobiology, University of Tennessee, Memphis 38104.
Wu
H
H
Byrne
M
M
Goldring
M B
MB
Seyer
J M
JM
Jaenisch
R
R
Krane
S M
SM
Mainardi
C L
CL
D000595:Amino Acid Sequence; D000818:Animals; D003094:Collagen / Q000235:genetics* / Q000378:metabolism; D017364:Collagenases / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D020784:Matrix Metalloproteinase 8; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009504:Neutrophils / Q000201:enzymology*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
D003094:Collagen; D017364:Collagenases; D020784:Matrix Metalloproteinase 8