Primary sequence and domain structure of chicken vinculin

摘要: Abstract We have determined the complete sequence of chick vinculin from two overlapping cDNA clones. The mRNA consists 262 bp 5' untranslated sequence, an open reading frame 3195 (excluding initiation codon) and a long 3' (greater than 2 kb). Chick contains 1066 amino acid residues, has deduced molecular mass 116,933 Da. Analysis domain structure shows that molecule can be cleaved by V8 proteinase into 90 kDa globular head 32 tail region, latter which could further 27 polypeptide. N-terminus vinculin, three 112-residue repeats (residues 259-589), extends to approximately residue 850. Gel overlay experiments show it also binding site for cytoskeletal protein talin. talin-binding was localized N-terminal 398 residues expression in vitro this region cloned Bluescript SK+ vector. domains are apparently separated proline-rich V8-proteinase-cleavage sites candidate tyrosine (822)-phosphorylation site. Secondary-structure prediction suggests contain alpha-helical regions short stretches turn/coil. Comparison with partial human reveals is highly conserved protein. In chickens Southern-blot analysis consistent single gene, therefore likely its higher-molecular-mass isoform termed metavinculin, arise through alternative splicing.