Biosynthesis, secretion and extracellular localization of anchorin CII, a collagen-binding protein of the calpactin family.

M. Pfäffle, F. Ruggiero, H. Hofmann, M. P. Fernández, O. Selmin, Y. Yamada, R. Garrone, K. von der Mark

Research output: Contribution to journalArticle

86 Scopus citations

Abstract

The amino acid sequence of anchorin CII, a collagen-binding protein isolated originally from chondrocyte membranes, was previously determined by sequencing of cDNA and proteolytic fragments of the protein. Computer analysis of the protein sequence revealed four internal repeats of approximately 70-80 residues, each containing a highly conserved consensus sequence of 17 residues. These repeats show considerable homology with sequences in human and bovine calpactin, lipocortin, endonexin and protein II, which are members of a family of Ca2+- and phospholipid-binding proteins, as well as major substrates of tyrosine kinases. While these proteins have been located at the inner side of the plasma membrane of fibroblasts and epithelial cells, here we present experimental evidence that anchorin CII is at least partially released from cells and binds to the outer cell surface. Biosynthesis studies in cell-free systems and in cell culture indicate that anchorin CII is not processed, which is consistent with the absence of signal sequences from the protein. Yet, pulse-chase experiments show that anchorin is released into the culture medium of fibroblasts after 30 min, and in chondrocyte cultures after 20 h. Anchorin CII was located to the outer cell surface of chondrocytes by lactoperoxidase-catalyzed cell surface iodination as well as by antibody labeling both at light- and electron-microscopical level. The pericellular localization of anchorin CII is consistent with the notion that this protein is involved in the interaction of chondrocytes and fibroblasts with extracellular collagen.

Original languageEnglish (US)
Pages (from-to)2335-2342
Number of pages8
JournalThe EMBO journal
Volume7
Issue number8
StatePublished - Aug 1988
Externally publishedYes

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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    Pfäffle, M., Ruggiero, F., Hofmann, H., Fernández, M. P., Selmin, O., Yamada, Y., Garrone, R., & von der Mark, K. (1988). Biosynthesis, secretion and extracellular localization of anchorin CII, a collagen-binding protein of the calpactin family. The EMBO journal, 7(8), 2335-2342.