Deletion of the pro-α1(I) N-propeptide affects secretion of type I collagen in Chinese hamster lung cells but not in Mov-13 mouse cells

S. T. Lee, S. Lee, D. P. Peters, G. G. Hoffman, A. Stacey, D. S. Greenspan

Research output: Contribution to journalArticle

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Abstract

We have introduced two mutations into a full-length human pro-α1(I) cDNA that delete 114 amino acids or the entire 139 amino acids of the N-propeptide domain. Wild-type and mutated versions of the cDNA were introduced into cultured Chinese hamster lung (CHL) cells, which do not produce endogenous type I collagen, and into Mov-13 mouse cells, which produce endogenous pro- α2(I) chains but not pro-α1(I) chains. As judged by resistance to proteases, neither mutation impaired intracellular triple helical assembly of human α1(I) homotrimers in CHL cells, or of chimeric type I collagen comprised of human α1(I) and mouse α2(I) chains in Mov-13 cells. Thus, the N-propeptide is not necessary for intracellular assembly of the main helical collagen domain of type I collagen. In CHL cells the rate of secretion of the mutant homotrimers was greatly reduced as compared to wild type homotrimers, and by immunofluorescence and immunoelectron microscopy, the mutant chains were shown to be accumulated in large vesicular expansions of the rough endoplasmic reticulum. When such cells were retransfected with cDNA encoding wild-type human α2(I) chains, mutant α1(I) chains were not rescued and heterotrimers containing the mutant chains were also retained in the intracellular vesicles. By contrast, deletion of the N-propeptide did not affect secretion of heterotrimers containing mutant chains from Mov-13 cells. Thus, an intact N-propeptide appears necessary for efficient secretion of type I collagen from some but not all cell types.

Original languageEnglish
Pages (from-to)24126-24133
Number of pages8
JournalJournal of Biological Chemistry
Volume267
Issue number33
Publication statusPublished - 1992 Dec 1

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Collagen Type I
Cricetulus
Lung
Complementary DNA
Amino Acids
Microscopic examination
Peptide Hydrolases
Collagen
Mutation
Immunoelectron Microscopy
Rough Endoplasmic Reticulum
Fluorescence Microscopy

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Lee, S. T. ; Lee, S. ; Peters, D. P. ; Hoffman, G. G. ; Stacey, A. ; Greenspan, D. S. / Deletion of the pro-α1(I) N-propeptide affects secretion of type I collagen in Chinese hamster lung cells but not in Mov-13 mouse cells. In: Journal of Biological Chemistry. 1992 ; Vol. 267, No. 33. pp. 24126-24133.
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abstract = "We have introduced two mutations into a full-length human pro-α1(I) cDNA that delete 114 amino acids or the entire 139 amino acids of the N-propeptide domain. Wild-type and mutated versions of the cDNA were introduced into cultured Chinese hamster lung (CHL) cells, which do not produce endogenous type I collagen, and into Mov-13 mouse cells, which produce endogenous pro- α2(I) chains but not pro-α1(I) chains. As judged by resistance to proteases, neither mutation impaired intracellular triple helical assembly of human α1(I) homotrimers in CHL cells, or of chimeric type I collagen comprised of human α1(I) and mouse α2(I) chains in Mov-13 cells. Thus, the N-propeptide is not necessary for intracellular assembly of the main helical collagen domain of type I collagen. In CHL cells the rate of secretion of the mutant homotrimers was greatly reduced as compared to wild type homotrimers, and by immunofluorescence and immunoelectron microscopy, the mutant chains were shown to be accumulated in large vesicular expansions of the rough endoplasmic reticulum. When such cells were retransfected with cDNA encoding wild-type human α2(I) chains, mutant α1(I) chains were not rescued and heterotrimers containing the mutant chains were also retained in the intracellular vesicles. By contrast, deletion of the N-propeptide did not affect secretion of heterotrimers containing mutant chains from Mov-13 cells. Thus, an intact N-propeptide appears necessary for efficient secretion of type I collagen from some but not all cell types.",
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Deletion of the pro-α1(I) N-propeptide affects secretion of type I collagen in Chinese hamster lung cells but not in Mov-13 mouse cells. / Lee, S. T.; Lee, S.; Peters, D. P.; Hoffman, G. G.; Stacey, A.; Greenspan, D. S.

In: Journal of Biological Chemistry, Vol. 267, No. 33, 01.12.1992, p. 24126-24133.

Research output: Contribution to journalArticle

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T1 - Deletion of the pro-α1(I) N-propeptide affects secretion of type I collagen in Chinese hamster lung cells but not in Mov-13 mouse cells

AU - Lee, S. T.

AU - Lee, S.

AU - Peters, D. P.

AU - Hoffman, G. G.

AU - Stacey, A.

AU - Greenspan, D. S.

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N2 - We have introduced two mutations into a full-length human pro-α1(I) cDNA that delete 114 amino acids or the entire 139 amino acids of the N-propeptide domain. Wild-type and mutated versions of the cDNA were introduced into cultured Chinese hamster lung (CHL) cells, which do not produce endogenous type I collagen, and into Mov-13 mouse cells, which produce endogenous pro- α2(I) chains but not pro-α1(I) chains. As judged by resistance to proteases, neither mutation impaired intracellular triple helical assembly of human α1(I) homotrimers in CHL cells, or of chimeric type I collagen comprised of human α1(I) and mouse α2(I) chains in Mov-13 cells. Thus, the N-propeptide is not necessary for intracellular assembly of the main helical collagen domain of type I collagen. In CHL cells the rate of secretion of the mutant homotrimers was greatly reduced as compared to wild type homotrimers, and by immunofluorescence and immunoelectron microscopy, the mutant chains were shown to be accumulated in large vesicular expansions of the rough endoplasmic reticulum. When such cells were retransfected with cDNA encoding wild-type human α2(I) chains, mutant α1(I) chains were not rescued and heterotrimers containing the mutant chains were also retained in the intracellular vesicles. By contrast, deletion of the N-propeptide did not affect secretion of heterotrimers containing mutant chains from Mov-13 cells. Thus, an intact N-propeptide appears necessary for efficient secretion of type I collagen from some but not all cell types.

AB - We have introduced two mutations into a full-length human pro-α1(I) cDNA that delete 114 amino acids or the entire 139 amino acids of the N-propeptide domain. Wild-type and mutated versions of the cDNA were introduced into cultured Chinese hamster lung (CHL) cells, which do not produce endogenous type I collagen, and into Mov-13 mouse cells, which produce endogenous pro- α2(I) chains but not pro-α1(I) chains. As judged by resistance to proteases, neither mutation impaired intracellular triple helical assembly of human α1(I) homotrimers in CHL cells, or of chimeric type I collagen comprised of human α1(I) and mouse α2(I) chains in Mov-13 cells. Thus, the N-propeptide is not necessary for intracellular assembly of the main helical collagen domain of type I collagen. In CHL cells the rate of secretion of the mutant homotrimers was greatly reduced as compared to wild type homotrimers, and by immunofluorescence and immunoelectron microscopy, the mutant chains were shown to be accumulated in large vesicular expansions of the rough endoplasmic reticulum. When such cells were retransfected with cDNA encoding wild-type human α2(I) chains, mutant α1(I) chains were not rescued and heterotrimers containing the mutant chains were also retained in the intracellular vesicles. By contrast, deletion of the N-propeptide did not affect secretion of heterotrimers containing mutant chains from Mov-13 cells. Thus, an intact N-propeptide appears necessary for efficient secretion of type I collagen from some but not all cell types.

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