Interactions between globular proteins and F-actin in isotonic saline solution.
Solutions of each of three different globular proteins (cytochrome c, chromophorically labeled serum albumin, and chromophorically labeled aldolase), mixed with another unlabeled globular protein or with fibrous actin, were prepared in pH 8.0 Tris-HCl buffer containing 0.15 M NaCl. Each solution was centrifuged at low speed, at 5 degrees C, until unassociated globular protein in solution achieved sedimentation equilibrium. Individual absorbance gradients of both macrosolutes in the mixtures subsequent to centrifugation were obtained via optical scans of the centrifuge tubes at two wavelengths. The gradients of each macrosolute in mixtures of two globular proteins revealed no association of globular proteins under the conditions of these experiments, but perturbation of the gradients of serum albumin, aldolase, and cytochrome c in the presence of F-actin indicated association of all three globular proteins with F-actin. Perturbation of actin gradients in the presence of serum albumin and aldolase suggested partial depolymerization of the F-actin by the globular protein. Analysis of the data with a simple phenomenological model relating free globular protein, bound globular protein, and total actin concentration provided estimates of the respective equilibrium constants for association of serum albumin and aldolase with F-actin, under the conditions of these experiments, of the order of 0.1 microM-1.
Lakatos
S
S
Laboratory of Biochemical Pharmacology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892.
Minton
A P
AP
D000199:Actins / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D002499:Centrifugation, Density Gradient; D003574:Cytochrome c Group / Q000378:metabolism*; D005634:Fructose-Bisphosphate Aldolase / Q000378:metabolism*; D008954:Models, Biological; D011817:Rabbits; D012710:Serum Albumin, Bovine / Q000378:metabolism*; D012965:Sodium Chloride; D012996:Solutions; D013056:Spectrophotometry, Ultraviolet
D000199:Actins; D003574:Cytochrome c Group; D012996:Solutions; D012710:Serum Albumin, Bovine; D012965:Sodium Chloride; D005634:Fructose-Bisphosphate Aldolase