Actin-fragmin interactions as revealed by chemical cross-linking.
A one to one complex of actin and fragmin (a capping protein from Physarum polycephalum plasmodia) was cross-linked with 1-ethyl-3-[3-(dimethylamino)propyl] carbodiimide. The cross-linking reaction generated two cross-linked products with slightly different molecular weights (88 000 and 90 000) as major species. They were cross-linked products of one actin and one fragmin. The cross-linking site of fragmin in the actin sequence was determined by peptide mappings [Sutoh, K. (1982) Biochemistry 21, 3654-3661] after partial chemical cleavages of cross-linked products with hydroxylamine. The results indicated that the N-terminal segment of actin spanning residues 1-12 participated in cross-linking with fragmin. The cross-linker used in this study covalently bridges lysine side chains and side chains of acidic residues when they are in direct contact. Therefore, it seems that acidic residues in the N-terminal segment of actin (Asp-1, Glu-2, Asp-3, Glu-4, and Asp-11), at least some of them, are in the binding site of fragmin. It has already been shown that the same acidic segment of actin is in the binding site of myosin or depactin (an actin-depolymerizing protein isolated from starfish oocytes). We suggest that the unusual amino acid sequence of the N-terminal segment of actin makes its N-terminal region a favorable anchoring site for various types of actin-binding proteins.
D051339:Actin Depolymerizing Factors; D000199:Actins / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003432:Cross-Linking Reagents; D051345:Destrin; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D019811:Hydroxylamine; D006898:Hydroxylamines; D046911:Macromolecular Substances; D008840:Microfilament Proteins / Q000378:metabolism*; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D010804:Physarum / Q000378:metabolism; D011485:Protein Binding
D051339:Actin Depolymerizing Factors; D000199:Actins; D003432:Cross-Linking Reagents; D051345:Destrin; D006898:Hydroxylamines; D046911:Macromolecular Substances; D008840:Microfilament Proteins; D010446:Peptide Fragments; D019811:Hydroxylamine