Actin in the ciliated protozoan Climacostomum virens: purification by DNAse I affinity chromatography, electrophoretic characterization, and immunological analysis.
The anti-actin monoclonal antibody (mab) JLA20 (Lin: Proc. Natl. Acad. Sci. U.S.A. 78:2335-2339, 1981) labels a 43 kD protein on Western blots of Climacostomum cell extracts; this protein does not react with an anti-alpha-smooth muscle actin mab (Skalli et al.: J. Cell Biol. 103:2787-2796, 1986) nor with an anti-alpha-sarcomeric actin mab (Skalli et al.: Am. J. Pathol. 130:515-531, 1988). This protein binds to DNAse I and can be purified by DNAse I affinity chromatography. The affinity-purified actin also reacts with mab JLA20. Two-dimensional gel analysis reveals that Climacostomum actin focuses as three spots which are more basic than the mammalian actin isoforms. After addition of KCl, the affinity-purified actin polymerizes into filaments as shown by electron microscopy after negative staining.
Fahrni
J F
JF
Department of Zoology and Animal Biology, Sciences III, Geneva, Switzerland.
D000199:Actins / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D002846:Chromatography, Affinity; D003850:Deoxyribonuclease I; D004591:Electrophoresis, Polyacrylamide Gel; D056890:Eukaryota / Q000737:chemistry*; D015151:Immunoblotting; D008969:Molecular Sequence Data; D016415:Sequence Alignment
D000199:Actins; D000911:Antibodies, Monoclonal; D003850:Deoxyribonuclease I