Identification and partial purification of a large, variant form of type XII collagen.
A large, alternate form of type XII collagen has been identified in cultures of the human epidermoid cell line WISH. This form, designated XIIA, is comprised of alpha chains that are approximately 90 kDa larger than the 220-kDa alpha chain previously characterized in extracts of fetal chicken and bovine tissues. Results from both collagenase digestion and rotary shadow analysis of partially purified material show that the increase is due to a larger NC3 domain. While both the large (XIIA) and the small (XIIB) forms of type XII collagen are identified in pulse-chase radiolabeling of fetal bovine skin explant culture, they are not related in a precursor-product fashion. Inhibition studies with alpha, alpha'-dipyridyl indicate that proper folding of the collagen helix is required for complete assembly and secretion of type XIIA in WISH cell culture. The 310-kDa alpha 1A chain is likely to represent the bovine equivalent of a second translation product, estimated to be 340 kDa, predicted from analysis of one complete chick cDNA sequence. Additionally, the amino-terminal amino acid sequence of the 220-kDa bovine alpha 1B chain was determined. This sequence is very near a potential alternate splice site predicted from analysis of chicken type XII cDNA.
Lunstrum
G P
GP
Shriners Hospital for Crippled Children, Portland, Oregon.
McDonough
A M
AM
Marinkovich
M P
MP
Keene
D R
DR
Morris
N P
NP
Burgeson
R E
RE
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002461:Cell Line, Transformed; D002645:Chickens; D002852:Chromatography, Ion Exchange; D003094:Collagen / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D004247:DNA; D004591:Electrophoresis, Polyacrylamide Gel; D000078404:Epidermal Cells; D004817:Epidermis / Q000737:chemistry; D006801:Humans; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D017510:Protein Folding
D003094:Collagen; D004247:DNA