An actin-depolymerizing protein in embryonic chicken skeletal muscle: purification and characterization.
In embryonic skeletal muscle, a large amount of non-polymerized actin exists in the cytoplasm (Shimizu and Obinata [1986] J. Biochem. 99, 751-759). A 19-kDa protein (called 19K protein) which binds to G-actin was purified by sequential chromatography on DNase I-agarose, hydroxylapatite, SP-Sephadex, and Sephadex G-75, from the sarcoplasmic fraction of embryonic chicken skeletal muscle. This protein decreased the extent of actin polymerization at a steady state and increased the monomeric actin in a concentration-dependent fashion; it also caused quick depolymerization of F-actin, as determined by spectrophotometry at 237 nm, viscometry, DNase I inhibition assay, and electron microscopy. The molar ratio of 19K protein and actin interacting with each other was estimated to be 1:1. From these results, 19K protein was regarded as being actin depolymerizing protein. The amount of 19K protein in muscle decreased during development. The inhibitory action of 19K protein was removed by myosin or heavy meromyosin, and actin filaments were formed on the surface of myosin filaments when myosin filaments were added to a mixture of actin and 19K protein in a physiological salt solution. We propose that actin assembly is dually controlled in the developing muscle by the inhibitor(s) and an accelerator (myosin); this mechanism may enable the ordered assembly of actin and myosin in the early phase of myofibrillogenesis.
Graduate School of Science and Technology, Chiba University.
H
H
Abe
T
T
Obinata
D008841:Actin Cytoskeleton / Q000378:metabolism; D051339:Actin Depolymerizing Factors; D000199:Actins / Q000145:classification; D000818:Animals; D001923:Brain Chemistry; D055598:Chemical Phenomena; D002621:Chemistry; D002642:Chick Embryo; D051345:Destrin; D008840:Microfilament Proteins / Q000032:analysis / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D009132:Muscles / Q000032:analysis* / Q000166:cytology / Q000196:embryology; D009218:Myosins / Q000494:pharmacology; D011485:Protein Binding; D014783:Viscosity
D051339:Actin Depolymerizing Factors; D000199:Actins; D051345:Destrin; D008840:Microfilament Proteins; D009218:Myosins