title: Plasma actin depolymerizing factor has both calcium-dependent and calcium-independent effects on actin.
abstract: The effects of pig plasma actin depolymerizing factor (ADF) on both G-actin polymerization and F-actin fragmentation have been examined by using rabbit skeletal muscle actin labeled with N-(1-pyrenyl)iodoacetamide, a sensitive fluorescent probe for monomer to filament interconversion. Fluorescence data have been compared with results obtained by viscometry and by difference absorption measurements at 232 nm. Plasma ADF nucleates actin filament assembly in a Ca2+-dependent manner; actin polymerization rates are enhanced at greater than 10(-6) M Ca2+. The calcium concentration dependence of this effect, showing a shift in ADF nucleating capacity between 10(-6) and 10(-7) M Ca2+, is that expected for an intracellular regulatory effect, but in plasma, the protein would always be saturated with Ca2+. Although the rate of polymerization is markedly enhanced in the presence of calcium ions, the extent of polymerization (as determined by the amplitude of the fluorescence change or the specific viscosity) is reduced in the presence of ADF and shows little or no Ca2+ dependence. The critical concentration of actin monomers is increased in the presence of ADF whether calcium is present or not. When ADF is added to F-actin, there is an immediate fall in fluorescence. This conversion of filaments to monomers by ADF (as defined by the fluorescence changes) is unaffected by calcium concentration. Electron micrographs of F-actin treated with ADF show that the filaments are indeed shortened at both high and low calcium concentrations. Taken together, these observations are interpreted in terms of a model in which ADF has both Ca2+-sensitive and Ca2+-insensitive binding sites for actin.
mesh_terms: D051339:Actin Depolymerizing Factors; D000199:Actins / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D051345:Destrin; D006801:Humans; D007700:Kinetics; D046911:Macromolecular Substances; D008840:Microfilament Proteins; D009132:Muscles / Q000378:metabolism; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D011817:Rabbits; D013552:Swine
chemical_list: D051339:Actin Depolymerizing Factors; D000199:Actins; C495824:DSTN protein, human; D051345:Destrin; D046911:Macromolecular Substances; D008840:Microfilament Proteins; D011506:Proteins; D002118:Calcium