A gelsolin-related actin-severing protein with fully reversible actin-binding properties from the tail muscle of crayfish, Astacus leptodactylus.
A Ca(2+)-dependent actin-severing protein was purified from the tail muscle of the crayfish Astacus leptodactylus. The isolation procedure involved extraction at low ionic strength in the presence of EGTA, followed by ammonium sulfate fractionation, ion-exchange chromatography and gel filtration. The purified crayfish actin modulator appeared as a single band with a molecular mass of 105 kDa on SDS/PAGE. The crustacean actin modulator revealed basic functional properties in common with vertebrate gelsolin, like the Ca(2+)-activated severing of F-actin and the nucleation of actin polymerization. However, both proteins differed in major aspects: Ca2+ activation of crayfish actin modulator started at lower threshold concentrations (0.1 microM). The effect of the modulator on shortening the nucleation phase of actin polymerization was significantly weaker at lower modulator/actin ratios. The modulator formed three distinct stoichiometric complexes with G-actin, identified as binary, ternary and quaternary. Binding of G-actin occurred in a low cooperative manner and was completely reversible by EGTA. Despite some properties being similar to those of villin, crayfish actin modulator did not cross-link actin filaments. It is regarded in principle as a gelsolin-type protein, but with characteristic functional deviations from vertebrate gelsolin.
Bock
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Institut für Zoomorphologie und Zellbiologie, Heinrich-Heine-Universität Düsseldorf, Germany.
Hinssen
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D'Haese
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D000199:Actins / Q000378:metabolism; D000818:Animals; D003400:Astacoidea; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003598:Cytoskeletal Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D018260:Gelsolin; D008970:Molecular Weight; D009124:Muscle Proteins / Q000302:isolation & purification*; D009132:Muscles / Q000737:chemistry*; D011506:Proteins / Q000302:isolation & purification*; D013623:Tail
D000199:Actins; D003598:Cytoskeletal Proteins; D018260:Gelsolin; D009124:Muscle Proteins; D011506:Proteins; C028514:actin-modulating proteins