Biosynthesis and characterization of type X collagen in human fetal epiphyseal growth plate cartilage.
We examined in vitro collagen biosynthesis by organ cultures from human fetal epiphyseal growth plate cartilage. The biosynthetic products were characterized by NaCl fractional precipitation, limited proteolytic digestion, and sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis. Organ cultures of human fetal epiphyseal growth plate cartilage synthesized large amounts of type X collagen in addition to type II, type IX, and type XI collagens. The individual polypeptide chains of human type X collagen migrated with an apparent M(r) of 45 kDa after proteolytic digestion with pepsin. The migration pattern of these molecules did not change when examined under reducing and nonreducing conditions, indicating that they did not contain intrahelical disfulfide bonds. Comparison of the rates at type X collagen biosynthesis at weeks 20 and 24 of human fetal development showed a marked increase of 24 weeks. Northern hybridization analysis of total RNA from freshly isolated epiphyseal growth plate chondrocytes with a cDNA corresponding to the carboxyl terminus of human type X collagen indicated that the developmental increase of type X collagen production is determined by pre-translational mechanisms.
Reginato
A M
AM
Department of Medicine, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Sanz-Rodriguez
C
C
Jimenez
S A
SA
D001483:Base Sequence; D015152:Blotting, Northern; D002358:Cartilage, Articular / Q000196:embryology / Q000378:metabolism*; D003094:Collagen / Q000378:metabolism*; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D004838:Epiphyses / Q000196:embryology / Q000378:metabolism*; D005333:Fetus / Q000166:cytology / Q000378:metabolism*; D006132:Growth Plate / Q000166:cytology / Q000378:metabolism*; D006706:Homeostasis; D006801:Humans; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000235:genetics; D009924:Organ Culture Techniques; D012333:RNA, Messenger / Q000378:metabolism; D012995:Solubility
D018076:DNA, Complementary; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D003094:Collagen