Structure of equine type I and type II collagens.
Collagen type I was purified from equine skin and flexor tendon, and type II collagen was purified from equine articular cartilage. The proteoglycans in these tissues were extracted, using guanidine HCl; the collagens were solubilized, using pepsin digestion, then were selectively precipitated with NaCl. Gel electrophoresis indicated that the precipitates contained only type I or type II collagen. Amino acid analysis indicated that collagen constituted > 97% of the total protein in the precipitates. Hydroxylation of proline was 42.0 +/- 0.6% (mean +/- SEM) in alpha 1(I) and alpha 2(I), and was 48.1 +/- 1.3% in alpha 1(II) chains. The hydroxylation of lysine was 23.2 +/- 0.7% in alpha 1(I) and 34.1 +/- 0.9% in alpha 2(I) chains from tendon, and 49.6 +/- 4.3% in alpha 1(II) chains from cartilage. The cyanogen bromide (CB)-peptide patterns of chromatographically purified equine alpha 2(I) and alpha 1(II) chains were similar to those published previously for rat, bovine, and human alpha 2 and alpha 1 chains. However, the CB-peptide pattern of the equine alpha 1(I) chain resembled the guinea pig alpha 1(I) chain, which has no methionine between CB7 and CB6. Purified equine alpha 1(I)CB7,6 contained no methionine, methionine sulfoxide, or homoserine lactone. Mass of 42.26 kd was determined by use of mass spectrometry, and N-terminal sequence analysis established that the first 12 amino acids of this CB7,6 were identical to the sequence of human alpha 1(I)CB7.(ABSTRACT TRUNCATED AT 250 WORDS)
Todhunter
R J
RJ
Department of Pathology, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853.
Wootton
J A
JA
Lust
G
G
Minor
R R
RR
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002358:Cartilage, Articular / Q000737:chemistry*; D002845:Chromatography; D003094:Collagen / Q000737:chemistry* / Q000302:isolation & purification; D003488:Cyanogen Bromide; D004591:Electrophoresis, Polyacrylamide Gel; D019791:Guanidine; D006146:Guanidines; D006736:Horses; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D010449:Peptide Mapping; D012867:Skin / Q000737:chemistry*; D013710:Tendons / Q000737:chemistry*
D000596:Amino Acids; D006146:Guanidines; D010446:Peptide Fragments; D003094:Collagen; D019791:Guanidine; D003488:Cyanogen Bromide