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Role of the pro‐α2(I) COOH‐terminal region in assembly of type I collagen: Truncation of the last 10 amino acid residues of pro‐α2(I) chain prevents assembly of type I collagen heterotrimer
Author(s) -
Lim Ailee,
Doyle Sharon A.,
Balian Gary,
Smith Barbara D.
Publication year - 1998
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/(sici)1097-4644(19981101)71:2<216::aid-jcb7>3.0.co;2-y
Subject(s) - complementary dna , amino acid , type i collagen , immunoprecipitation , procollagen peptidase , trimer , microbiology and biotechnology , peptide sequence , transfection , messenger rna , chemistry , signal peptide , biochemistry , biology , gene , dimer , organic chemistry , endocrinology
Procollagen (Type I) contains a noncollagenous COOH‐terminal propeptide (C‐propeptide) hypothesized to be important in directing chain association and alignment during assembly. We previously expressed human pro‐α2(I) cDNA in rat liver epithelial cells, W8, that produce only pro‐α1(I) trimer collagen (Lim et al. [1994] Matrix Biol . 14: 21–30). In the resulting cell lines, α2(I) assembled with α1(I) forming heterotrimers. Using this cell system, we investigated the importance of the COOH‐terminal propeptide sequence of the pro‐α2(I) chain for normal assembly of type I collagen. Full‐length human pro‐α2(I) cDNA was cloned into expression vectors with a premature stop signal eliminating the final 10 amino acids. No triple‐helical molecules containing α2(I) were detected in transfected W8 cells, although pro‐α2(I) mRNA was detected. Additional protein analysis demonstrated that these cells synthesize small amounts of truncated pro‐α2(I) chains detected by immunoprecipitation with a pro‐α2(I) antibody. In addition, since the human‐rat collagen was less thermostable than normal intraspecies collagen, wild‐type and C‐terminal truncated mouse cDNAs were expressed in mouse D2 cells, which produced only type I trimers. Results from both systems were consistent, suggesting that the last 10 amino acid residues of the pro‐α2(I) chain are important for formation of stable type I collagen. J. Cell. Biochem. 71:216–232, 1998. © 1998 Wiley‐Liss, Inc.