An 18K protein from ascites hepatoma cell depolymerizes actin filaments rapidly.
Several actin binding proteins were isolated from ascites hepatoma cells AH7974 by DNase I affinity chromatography. Among them, a protein having a molecular weight of 18,000 was further purified by DEAE cellulose and hydroxyapatite column chromatographies and gel filtration on a Sephadex G-75 column. The 18K protein not only inhibits actin polymerization but also depolymerizes actin filaments. This conclusion was supported by viscosity and fluorescence intensity measurements and the DNase I inhibition assay. A chemical cross-linking experiment suggested that the 18K protein binds to monomeric actin and forms and 18K-actin 1:1 complex. The net depolymerization rate by the 18K protein measured by the DNase I inhibition assay was slower than the rapid reduction of the fluorescence intensity of pyrene-labeled F-actin upon addition of the 18K protein. This result suggests that the 18K protein not only binds to monomeric actin but also binds to actin filaments directly. The sedimentation assay showed that a part of the 18K protein was cosedimented with actin filaments. Electron microscopic observations demonstrated that the 18K protein decreased the amount of actin filaments and the remaining filaments appeared to be decorated and distorted by the 18K protein. The 18K protein had no Ca2+ ion sensitivity and exhibited the same effect on both this tumor actin and muscle actin.
D000199:Actins / Q000378:metabolism*; D000818:Animals; D001704:Biopolymers; D002352:Carrier Proteins / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity; D003599:Cytoskeleton / Q000378:metabolism; D003850:Deoxyribonuclease I; D018260:Gelsolin; D008114:Liver Neoplasms, Experimental / Q000378:metabolism*; D008840:Microfilament Proteins; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000302:isolation & purification* / Q000378:metabolism; D051381:Rats; D013050:Spectrometry, Fluorescence
D000199:Actins; D001704:Biopolymers; D002352:Carrier Proteins; D018260:Gelsolin; D008840:Microfilament Proteins; D009363:Neoplasm Proteins; C016953:brevin; D003850:Deoxyribonuclease I