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Human plasma actin-depolymerizing factor. Purification, biological activity and localization in leukocytes and platelets

Patebex, P.
Published in European Journal of Biochemistry. 1985, vol. 146, no. 2, p. 267-276
Abstract The plasma and serum of humans and various animal species exert an actin-depolymerizing activity. Human actin-depolymerizing factor (ADF) has been purified by ammonium sulfate fractionation, DEAE-cellulose and blue-Sepharose chromatography. It is a single polypeptide of approximately 90 kDa, with a pI between 6.0 and 6.5. ADF is heat and trypsin-sensitive, inactivated by EGTA, not stained by HIO4/Schiff on sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS/PAGE), and not retained on a concanavalin-A-Sepharose column. Incubation of ethanol-fixed cultured cells or unfixed cryostat tissue sections with ADF abolishes immunofluorescent actin staining, by a mechanism which involves extraction of actin from the preparations. ADF promotes fragmentation and depolymerization of actin filaments as shown by electron microscopy, differential ultracentrifugation and DNAse I inhibition assay. This depolymerized actin retains its mobility on SDS/PAGE and is able to repolymerize in the presence of EGTA. Human white blood cells and platelets (but neither human fibroblasts nor white blood cells and platelets from pig, rat and rabbit) contain a 90-kDa protein reacting with an antibody raised in rabbit against human ADF as judged by immunofluorescence and immunoblotting techniques. Immunoblots of human granulocyte subcellular fractions suggest that the protein reacting with ADF antibody is present in the soluble cytoplasmic fraction. ADF may play a role in solubilization of plasma actin and in the intracellular organization of actin, and should be useful for the evaluation of the relative stability of cytoplasmic actin filaments in various physiological and pathological processes.
Keywords Actin Depolymerizing FactorsAntibody FormationBlood Platelets/ metabolismBlood ProteinsCapZ Actin Capping ProteinCells, CulturedDestrinFluorescent Antibody TechniqueFreezingHumansLeukocytes/ metabolismMicrofilament ProteinsMuscles/drug effects/ultrastructureNeutrophils/metabolismProteins/immunology/pharmacologyStaining and LabelingUltracentrifugation
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PMID: 2578390
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