Purification and properties of bovine nasal hyaline cartilage collagenase

Int J Biochem. 1990;22(11):1273-82. doi: 10.1016/0020-711x(90)90309-q.

Abstract

1. Collagenase from bovine nasal hyaline cartilage was extracted with 1 and 3 M NaCl in Tris-CaCl2 buffer. 2. Two peaks of collagenase activity were revealed on DE52 ion exchange column, collagenase 1 and collagenase 2. 3. The apparent mol. wt of collagenase 1 and 2 as determined by SDS-PAGE were 68 and 43 kDa, respectively. 4. Both enzymes degrade native collagen type II into two characteristic products, TCA(3/4) and TCB(1/4), at 25 degrees C and pH 7.6. 5. Trypsin and aminophenylmercuric acetate were capable of increasing the collagenase 1 activity. 6. The two enzymes can be characterized as metalloproteinases since they were inhibited by EGTA and 1,10-phenanthroline. The use of proteinase inhibitors (N-ethylmaleimide, iodoacetic acid, phenylmethylsulphonyl fluoride, soybean trypsin inhibitor, pepstatin, dithiothreitol) showed that the enzymes do not contain serine, cysteine or aspartic acid in their active sites.

MeSH terms

  • Animals
  • Cartilage / enzymology*
  • Cattle
  • Chromatography, Gel
  • Collagen / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Hydrogen-Ion Concentration
  • In Vitro Techniques
  • Microbial Collagenase / chemistry
  • Microbial Collagenase / isolation & purification*
  • Microbial Collagenase / metabolism
  • Molecular Weight
  • Nose

Substances

  • Collagen
  • Microbial Collagenase