Type IX Collagen NC1 Domain Peptides Can Trimerize in Vitro without Forming a Triple Helix

• Published in: The Journal of biological chemistry Vol. 271; no. 23; pp. 13781 - 13785
• Main Authors: Mechling, Diane E., Gambee, Jay E., Morris, Nicholas P., Sakai, Lynn Y., Keene, Douglas R., Mayne, Richard, Bächinger, Hans Peter
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 07-06-1996; American Society for Biochemistry and Molecular Biology
• Subjects: Amino Acid Sequence; Animals; Antibodies, Monoclonal; Chickens; Collagen - chemistry; Collagen - genetics; Collagen - immunology; Disulfides - chemistry; Epitope Mapping; Humans; In Vitro Techniques; Mice; Molecular Sequence Data; Molecular Structure; Peptide Fragments - chemical synthesis; Peptide Fragments - chemistry; Peptide Fragments - genetics; Protein Conformation; Protein Structure, Secondary
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.271.23.13781

Synthetic peptides of the three chains of type IX collagen consisting of the carboxyl-terminal end of the COL1 domain and the complete NC1 domain were characterized by circular dichroism spectroscopy and analyzed for their ability to assemble into trimers. In vitro association and oxidation result in disulfide-linked oligomers as shown by molecular sieve chromatography and SDS-polyacrylamide electrophoresis. Whereas the individual peptides show a tendency to self-associate, when an equimolar amount of the three peptides was oxidized, a heterotrimer of the three chains was observed. This heterotrimer is recognized by a monoclonal antibody against the disulfide-linked NC1 domain of chicken type IX collagen, indicating the correct formation of the disulfide bonds. Circular dichroism measurements show that under the association conditions used, a triple helix does not form between the chains. These results indicate that these peptides contain all the necessary information for chain selection and assembly.