A neutral collagenase from human gastric mucosa.
Biopsy specimens of human gastric mucosa, maintained in culture for 7 days in the absence of serum, released a collagen-degrading enzyme into the medium. The yield of active enzyme reached a maximum after 2-3 days, and viable tissue, capable of protein synthesis, was essential for its production. 2. At 25 degrees C the enzyme attacked undenatured collagen in solution, resulting in a 55% loss of specific viscosity and producing the two products TCA and TCB characteristic of neutral-collagenase action. 3. Electron microscopy of segment-long-spacing crystallites of these reaction products showed the exact cleavage locus of the collagen molecules to be between bands 43 and 44 (I-43). The larger TCA and smaller TCB products were fragments representing 77 and 23% respectively of the length of the collagen molecule. 4. Optimal enzyme activity was observed over the pH range 7.5-8.5 and a mol.wt. of approx. 38000 was derived from gel-filtration studies. 5. The enzyme was shown to be inhibited by the human serum proteins alpha2-macroglobulin and a smaller component of mol.wt. approx. 40000; alpha1-anti-trypsin was not inhibitory. 6. EDTA, 1, 10-phenanthroline, cysteine and dithiothreitol all inhibited collagenase activity. 7. The gastric enzyme has properties similar to other well characterized collagenases, but differences exist with respect to its molecular size and the site of attack on the collagen molecule.
D001798:Blood Proteins / Q000032:analysis; D046508:Culture Techniques; D004492:Edetic Acid / Q000494:pharmacology; D004589:Electrophoresis, Disc; D005753:Gastric Mucosa / Q000201:enzymology*; D005947:Glucose / Q000032:analysis; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006909:Hydroxyproline / Q000032:analysis; D003012:Microbial Collagenase / Q000032:analysis* / Q000037:antagonists & inhibitors; D008854:Microscopy, Electron; D008970:Molecular Weight; D010618:Phenanthrolines / Q000494:pharmacology; D014783:Viscosity
D001798:Blood Proteins; D010618:Phenanthrolines; D004492:Edetic Acid; D003012:Microbial Collagenase; D005947:Glucose; D006909:Hydroxyproline