Thermal stability of human-fibroblast-collagenase-cleavage products of type-I and type-III collagens.
Rat skin type-I and type-III collagens were degraded by human fibroblast collagenase at a temperature below the 'melting' temperature for the two resulting fragments, namely the N-terminal three-fourths, TCA, and the C-terminal one-fourth, TCB. The specific cleavage of the collagen was confirmed by electrophoresis and determination of molecular length by electron microscopy. The two fragments were separated by gel filtration and the thermal stabilities of the isolated fragments were determined. For type-I collagen, the 'melting' temperatures of the two fragments were found to differ by only 0.5 degrees C and were 4.5-5.0 degrees C below that of the uncleaved molecule. The 'melting' temperatures of the uncleaved molecule and the N-terminal fragment were independent of the extent of N-terminal intramolecular cross-linking. For type-III collagen, the 'melting' temperatures of the fragments were found to differ by 1.3 degrees C. The small fragments of the two types of collagen 'melted' at the same temperature, whereas the large type-III fragment 'melted' at a slightly higher temperature than did the large type-I fragment. Reduction of the disulphide bonds located in the C-terminal type-III fragment did not affect the thermal stability of this fragment. The thermal stability of uncleaved type-III collagen was found to be variable, but the reason for this is not known at present.
Danielsen
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CC
Department of Connective Tissue Biology, University of Aarhus, Denmark.
D000818:Animals; D002850:Chromatography, Gel; D003094:Collagen; D004355:Drug Stability; D005347:Fibroblasts / Q000201:enzymology; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008297:Male; D003012:Microbial Collagenase / Q000378:metabolism*; D008854:Microscopy, Electron; D010446:Peptide Fragments / Q000032:analysis; D011489:Protein Denaturation; D051381:Rats; D011919:Rats, Inbred Strains
D010446:Peptide Fragments; D003094:Collagen; D003012:Microbial Collagenase