title: Alternative splicing of the first F3 domain from chicken collagen XIV affects cell adhesion and heparin binding.
abstract: The N terminus of chicken collagen XIV is subject to alternative splicing. The longer isoform contains a fibronectin type III (F3) domain at its N terminus, whereas the shorter isoform is lacking this domain. Alternative splicing of the F3 domain is developmentally regulated. At early embryonic stages, both isoforms are expressed, whereas after hatching only the longer isoform is expressed. When immobilized on plastic dishes, the recombinant F3 domain promotes the adhesion of mesenchymal cells. Attachment to this domain is specifically inhibited by heparin but not by other glycosaminoglycans. Molecular modeling studies illustrate that the first F3 domain harbors a positively charged groove, which may accommodate the negatively charged heparin chain. Site-directed mutagenesis of a single lysine residue within this groove abolishes the cell binding activity but does not affect the heparin binding activity. Cell binding and heparin binding are therefore two functionally distinct properties shared by the N-terminal F3 domain. When full-length collagen XIV polypeptides that either contain or lack the first F3 domain are tested on heparin-Sepharose, a pronounced difference in their relative affinity is observed. Thus, alternative splicing of the N-terminal F3 domain influences the interaction of this FACIT (fibril-associated collagens with interrupted triple helices) collagen with cells and with glycosaminoglycans.
authors.affiliation: M. E. Müller Institute, University of Bern, CH-3010 Bern, Switzerland.
authors.forename: M
authors.initials: M
authors.lastname: Imhof
authors.forename: B
authors.initials: B
authors.lastname: Trueb
mesh_terms: D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002448:Cell Adhesion / Q000502:physiology*; D002460:Cell Line; D002645:Chickens; D003094:Collagen / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D017931:DNA Primers; D018076:DNA, Complementary; D008958:Models, Molecular; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D020033:Protein Isoforms; D011993:Recombinant Fusion Proteins; D003094:Collagen