Isolation of human type X collagen and immunolocalization in fetal human cartilage.
Type X collagen was extracted with 1 M NaCl and 10 mM dithiothreitol at neutral pH from fetal human growth plate cartilage and purified to homogeneity by gel filtration and anion-exchange chromatography. The purified protein migrates in SDS/polyacrylamide gels with an apparent Mr of 66,000 under reducing conditions, and as a high-Mr oligomer under non-reducing conditions. Purified collagenase digests most of the molecule; pepsin digestion at 4 degrees C decreases the Mr of the monomer to 53,000. A rabbit antiserum was raised against purified human type X collagen; the IgG fraction was specific for this collagen by criteria of ELISA and immunoblotting after absorption with collagen types I, II, VI, IX and XI. Immunohistological studies localized type X collagen exclusively in the zone of hypertrophic and calcifying cartilage.
Kirsch
T
T
Clinical Research Units for Rheumatology, University of Erlangen-Nürnberg, Federal Republic of Germany.
von der Mark
K
K
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002356:Cartilage / Q000737:chemistry*; D003094:Collagen / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D005333:Fetus / Q000737:chemistry*; D006651:Histocytochemistry; D006801:Humans; D008970:Molecular Weight; D011817:Rabbits
D000596:Amino Acids; D003094:Collagen