Isolation and sequence analysis of the glycosaminoglycan attachment site of type IX collagen.
Type IX collagen from chick embryonic cartilage is unique among the collagens in that it contains chondroitin sulfate covalently linked to the alpha 2(IX) polypeptide chain. We have isolated and sequenced the glycosaminoglycan-containing peptide released by collagenase digestion from type IX collagen, labeled biosynthetically with [35SO4] and 3H-aminoacids. This peptide was purified by gel filtration and, following chondroitinase ABC digestion, by reverse-phase high performance liquid chromatography. The amino acid sequence obtained for this peptide has 23 residues, beginning and ending with a collagenous sequence, indicating that it spans an internal noncollagenous domain. Comparison of this sequence with the one predicted from cDNA clone pYN 1738 for the alpha 1(IX)chain and pYN 1731 and pDM 222 for the alpha 2(IX)chain revealed the peptide to be the noncollagenous NC3 domain of alpha 2(IX). The glycosylated sequence Val-Glu-Gly-Ser*-Ala-Asp- of type IX collagen does not have the Ser-Gly normally functioning as the attachment sequence but does have an acidic residue preceding the serine which should improve the acceptability of this sequence for the xylosyltransferase. That it is an adequate acceptor can be inferred from the observation that type IX collagen carries a glycosaminoglycan chain on over 70% of the molecules isolated.
Huber
S
S
Laboratorium für Biochemie I, Eidgenössische Technische Hochschule, Zürich, Switzerland.
Winterhalter
K H
KH
Vaughan
L
L
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002356:Cartilage / Q000032:analysis; D002500:Centrifugation, Isopycnic; D002642:Chick Embryo; D002808:Chondroitin Lyases / Q000378:metabolism; D003094:Collagen / Q000032:analysis* / Q000378:metabolism; D006025:Glycosaminoglycans / Q000378:metabolism*; D010446:Peptide Fragments / Q000032:analysis / Q000302:isolation & purification*
D006025:Glycosaminoglycans; D010446:Peptide Fragments; D003094:Collagen; D002808:Chondroitin Lyases