Incomplete processing of type II procollagen by a rat chondrosarcoma cell line.
The Swarm rat chondrosarcoma cell line, RCS-LTC, deposits an extracellular matrix that contains the typical type II, IX, and XI collagen phenotype of hyaline cartilage, but the fibrils appear abnormally thin. By N-terminal sequence analysis, the type II collagen from the matrix was shown to have retained its N-propeptides with no evidence of normal processing to type II collagen. Amplification and sequencing of cDNA prepared from the pro alpha1(II) mRNA of these cells showed a normal N-propeptide cleavage site. Furthermore, the type II N-procollagen could be processed to type II collagen by incubation with culture medium from normal chondrocytes. The findings indicate that the RCS-LTC cell line fails to express an active type II procollagen N-proteinase and, therefore, offers a useful culture system in which to study the role of N-propeptide removal in fibrillogenesis.
Fernandes
R J
RJ
Department of Orthopaedics, University of Washington, Seattle 98195, USA. rjf@u.washington.edu
Schmid
T M
TM
Harkey
M A
MA
Eyre
D R
DR
D000595:Amino Acid Sequence; D000818:Animals; D002813:Chondrosarcoma / Q000378:metabolism*; D003094:Collagen / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D018076:DNA, Complementary; D005091:Exons; D007700:Kinetics; D008969:Molecular Sequence Data; D011347:Procollagen / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D011348:Procollagen N-Endopeptidase / Q000172:deficiency / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; D011347:Procollagen; D012333:RNA, Messenger; D011994:Recombinant Proteins; D003094:Collagen; D011348:Procollagen N-Endopeptidase