Several peptides were isolated from tryptic digests of insoluble calf aorta matrix by chromatography. Reductive pyridylethylation of a tryptic 15 kDa pool released fragments deriving from the C-terminus of type III collagen. A 50-residue peptide TC(III) was shown by sequence analysis to be the C-terminal peptide from the α1(III)-chain, containing a helical and non-helical region of equal sizes. The peptide was further digested with collagenase to give ColC(III), comprising the complete C-terminal non-helical region of α1(III) including a hydroxylysine in position 16C. The peptide TC(III)×TN(III) was isolated, demonstrating covalent cross-linking between the C-terminal non-helical region of one type III molecule and the N-terminal helical cross-linking region of another. Its digestion with cyanogen bromide yielded the small fragments α1(III)CB3B* and α1(III)CB3C, confirming TN(III) as an N-terminal helical cross-link site. Sequence analysis of both TC(III)×TN(III) and its collagenase-derived cross-linked peptide ColC(III)×TN(III) established the 4D-staggered alignment of adjacent collagen III molecules. The cross-link structure of both peptides was mainly dihydroxylysinonorleucine with a small amount of hydroxylysinonorleucine, indicating that the lysine residues involved in formation of the cross-links are both hydroxylated. No pyridinoline or histidinohydroxylysinonorleucine cross-links were found within the non-reduced C-telopeptide region of type III collagen.
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September 1996
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Research Article|
September 01 1996
Cross-link analysis of the C-telopeptide domain from type III collagen
W. HENKEL
W. HENKEL
1Institut für Arterioskleroseforschung an der Universität Münster, Domagkstrasse 3, D-48149 Münster, Federal Republic of Germany
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Publisher: Portland Press Ltd
Received:
April 04 1996
Accepted:
May 02 1996
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1996
1996
Biochem J (1996) 318 (2): 497–503.
Article history
Received:
April 04 1996
Accepted:
May 02 1996
Citation
W. HENKEL; Cross-link analysis of the C-telopeptide domain from type III collagen. Biochem J 1 September 1996; 318 (2): 497–503. doi: https://doi.org/10.1042/bj3180497
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