Stepwise activation mechanisms of the precursors of matrix metalloproteinases 1 (tissue collagenase) and 3 (stromelysin).
Two zymogens of matrix metalloproteinases (MMPs), proMMP-1 (tissue procollagenase) and proMMP-3 (prostromelysin) were isolated from the culture medium of human rheumatoid synovial fibroplasts and their activation mechanisms by proteinases and 4-aminophenylmercuric acetate (APMA) were studied by kinetic and sequence analyses. Both zymogens were activated by unique stepwise activation mechanisms through which sequential processing events occur in the propeptide regions. The initial cleavage sites attacked by activator proteinases are located in the middle of the propeptides at Glu33-Lys-Arg-Arg-Asn37 in proMMP-1 and Phe34-Val-Arg-Arg-Lys-Asp39 in proMMP-3. The initial products of proMMP-1 generated by proteinases then undergo further autocleavage of the Thr64-Leu65 bond. The treatment of proMMP-1 and proMMP-3 with APMA results in the intramolecular cleavage of the Val67-Met68 and Glu68-Val69 bonds, respectively. The removal of a portion of propeptides results in conformational changes around the Gln80-Phe81 and His82-Phe83 bonds in respective intermediates of MMP-1 and MMP-3 and render them to rapid specific cleavage by MMP-3 to generate stable, fully active enzymes.
Nagase
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Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City 66103.
Suzuki
K
K
Enghild
J J
JJ
Salvesen
G
G
D000595:Amino Acid Sequence; D001665:Binding Sites; D017364:Collagenases; D003238:Connective Tissue / Q000201:enzymology; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D020781:Matrix Metalloproteinase 1; D008666:Metalloendopeptidases / Q000378:metabolism*; D003012:Microbial Collagenase / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data
D004792:Enzyme Precursors; D017364:Collagenases; D008666:Metalloendopeptidases; C022440:procollagenase; C048047:prostromelysin; D003012:Microbial Collagenase; D020781:Matrix Metalloproteinase 1