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 con...

Full description

Saved in:
Bibliographic Details
Published in:Biochemical journal 1987-11, Vol.247 (3), p.725-729
Main Author: Danielsen, C C
Format: Article
Language:English
Subjects:
Animals
Chromatography, Gel
Collagen
collagens
Drug Stability
Fibroblasts - enzymology
Hot Temperature
Hydrogen-Ion Concentration
Male
Microbial Collagenase - metabolism
Microscopy, Electron
Peptide Fragments - analysis
Protein Denaturation
Rats
Rats, Inbred Strains
thermal stability
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:0264-6021
1470-8728
DOI:10.1042/bj2470725