title: Mutation analysis of cadherin-4 reveals amino acid residues of EC1 important for the structure and function.
abstract: To clarify the structural basis of the cell adhesion activity of cadherins, we examined the effects of point mutations of well-conserved amino acid residues in the extracellular domain 1 of cadherin-4 (Cdh4) on the adhesion properties by alanine scanning mutagenesis. Mutations of two well-conserved aromatic amino acid residues in the extracellular domain 1 resulted in abnormal processing of Cdh4 molecules and no cell adhesion activity, whereas mutations of the corresponding aromatic amino acids in the extracellular domain 2 did not show these effects, suggesting a role for the two residues in the extracellular domain 1 in the folding and/or intracellular transport processes of Cdh4. Mutations of the amino acid residues suspected to be involved in strand dimer formation resulted in loss or significant decrease in cell adhesion activity. The mutant Cdh4s showed weak concentration at cell-cell adhesion sites and chemical cross-linking suggested that the strand dimer formation was actually impaired in the mutants. These results are consistent with the zipper model, in which the extracellular domain 1 of Cdh4 has intrinsic strand dimer formation activity in addition to adhesion dimer formation activity, both of which are involved in cell adhesion activity. The zipper model, however, needs further improvement to fully account for the present results.
authors.affiliation: Department of Clinical Research, Okura National Hospital, Okura, Tokyo, 157-8535, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Kitagawa
authors.forename: M
authors.initials: M
authors.lastname: Natori
authors.forename: S
authors.initials: S
authors.lastname: Murase
authors.forename: S
authors.initials: S
authors.lastname: Hirano
authors.forename: S
authors.initials: S
authors.lastname: Taketani
authors.forename: S T
authors.initials: ST
authors.lastname: Suzuki
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D015820:Cadherins / Q000737:chemistry / Q000235:genetics*; D002448:Cell Adhesion / Q000235:genetics; D002449:Cell Aggregation / Q000235:genetics; D017124:Conserved Sequence; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004252:DNA Mutational Analysis; D006801:Humans; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D016296:Mutagenesis; D017354:Point Mutation; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D014162:Transfection
chemical_list: D000596:Amino Acids; D015820:Cadherins; D003432:Cross-Linking Reagents; C083970:R-cadherin