Sequence of canine COL1A2 cDNA: nucleotide substitutions affecting the cyanogen bromide peptide map of the alpha 2(I) chain.
The alpha2 chain of canine type I collagen was characterized with both sequence analysis of COL1A2 cDNA and chemical analysis of alpha2(I) chains. The complete sequence of canine COL1A2 cDNA was determined from reverse-transcribed and polymerase chain reaction-amplified total RNA from cultured skin fibroblasts. Pepsin-digested and cyanogen bromide-digested type I collagen peptides were analyzed with chromatography, gel electrophoresis, and mass spectrometry. Identity between the sequences of canine and human COL1A2 cDNA was 90.9%, predicting conservation of the 3 cysteine residues required for C-propeptide registration and of cleavage sites for signal peptidase, procollagen N-proteinase, vertebrate collagenase, and procollagen C-proteinase. Conservation of all 50 lysine residues was also predicted, including preservation of the 1:2 asymmetry in the X:Y distribution of the 31 lysine residues in the alpha2(I) triple helix. The human and canine sequences differed in the location of Y-position proline residues and the presence of two unique canine cyanogen bromide peptides, alpha2 CB3b and alpha2 CB3c,5. Knowledge of the conserved and unique features of canine COL1A2 will be valuable for analysis of the expression, synthesis, and structure of type I collagen as well as studies of canine osteogenesis imperfecta.
College of Veterinary Medicine, Cornell University, Ithaca, New York, 14853, USA. bgc1@cornell.edu
B G
BG
Campbell
J A
JA
Wootton
J N
JN
MacLeod
R R
RR
Minor
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry* / Q000235:genetics*; D017124:Conserved Sequence; D003488:Cyanogen Bromide / Q000737:chemistry*; D018076:DNA, Complementary / Q000737:chemistry* / Q000235:genetics; D004285:Dogs; D005347:Fibroblasts; D006801:Humans; D008239:Lysine / Q000737:chemistry; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D010449:Peptide Mapping; D017354:Point Mutation / Q000235:genetics*; D011392:Proline / Q000737:chemistry
D018076:DNA, Complementary; D010446:Peptide Fragments; D003094:Collagen; D011392:Proline; D008239:Lysine; D003488:Cyanogen Bromide