Distribution of two alternatively spliced variants of the type II collagen N-propeptide compared with the C-propeptide in bovine chondrocyte pellet cultures.
We have analyzed the distribution of type II collagen N- and C-propeptides in the cell layers and culture medium of bovine articular chondrocyte pellet cultures. Two splice variants of the type II collagen N-propeptide were detected by immunoblotting and immunoassay, using a new anti-peptide antibody, while the C-propeptide was detected using a monoclonal antibody. Type II collagen molecules containing the N-propeptide were detected weakly in cell layers, but not in tissue culture medium of chondrocyte pellet cultures, and both splice variants were observed. Free N-propeptide could not be detected in cell layers or medium. Type II procollagen molecules containing the C-propeptide were detected strongly in cell layers, but not in tissue culture medium, while the free C-propeptide was detected in both cell layers and medium. Since the N- and C-propeptides must be synthesized in a 1:1 molar ratio, we conclude that the N-propeptide is metabolized more quickly than the C-propeptide in this system. Our model can be used to study regulation of procollagen synthesis and propeptidase activity.
Rebuck
N
N
Human Metabolism and Clinical Biochemistry in the Division of Biochemical and Musculoskeletal Medicine, University of Sheffield Medical School, Sheffield, S10 2RX, UK.
Croucher
L J
LJ
Hollander
A P
AP
D017398:Alternative Splicing / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D002135:Calcium-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D002358:Cartilage, Articular / Q000378:metabolism; D002417:Cattle; D019902:Chondrocytes / Q000378:metabolism*; D003094:Collagen / Q000235:genetics* / Q000378:metabolism; D024043:Collagen Type II; D017077:Culture Media, Conditioned / Q000737:chemistry; D000939:Epitopes; D005109:Extracellular Matrix / Q000378:metabolism; D007118:Immunoassay; D015151:Immunoblotting; D008969:Molecular Sequence Data; D011347:Procollagen / Q000378:metabolism; D011498:Protein Precursors / Q000235:genetics* / Q000378:metabolism; D014822:Vitreous Body
D000906:Antibodies; D002135:Calcium-Binding Proteins; D024043:Collagen Type II; D017077:Culture Media, Conditioned; D000939:Epitopes; D011347:Procollagen; D011498:Protein Precursors; C040072:chondrocalcin; D003094:Collagen