A cofilin-like protein is involved in the regulation of actin assembly in developing skeletal muscle.
An actin-binding protein of 20 kDa (called 20K protein) was purified from the sarcoplasmic fraction of embryonic chicken skeletal muscle. The properties of this protein were very similar to cofilin, which was discovered in porcine brain (Nishida et al. (1984) Biochemistry, 23, 5307-5313): it bound to both G- and F-actin, inhibited actin polymerization in a pH-dependent manner, inhibited binding of tropomyosin to F-actin, and had almost the same molecular size and pI as cofilin. A specific monoclonal antibody to 20K protein (MAB-22) was prepared to examine the expression and location of 20K protein during skeletal muscle development. When the whole protein lysates of embryonic and post-hatched chicken skeletal muscles were examined by means of immunoblotting combined with SDS-PAGE, 20K protein was detected in skeletal muscle through the developmental stages. Location of 20K protein in the cells differed between the embryonic and adult tissues; immunofluorescence staining of the cryosections of embryonic muscle with MAB-22 visualized irregular dot-like structures, but adult muscle sections were stained faintly and uniformly. 20K protein was present as a complex with actin in embryonic muscle, as judged by the ability to bind to a DNase I affinity column, while the same protein was free from actin in the cytoplasm of adult muscle. From these results, it is suggested that 20K protein regulates actin assembly transiently in developing skeletal muscle.
Abe
H
H
Graduate School of Science and Technology, Faculty of Science, Chiba University.
Ohshima
S
S
Obinata
T
T
D051339:Actin Depolymerizing Factors; D000199:Actins / Q000096:biosynthesis*; D000818:Animals; D002642:Chick Embryo; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D008840:Microfilament Proteins; D008970:Molecular Weight; D009124:Muscle Proteins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification; D009132:Muscles / Q000196:embryology / Q000378:metabolism*; D009218:Myosins / Q000502:physiology; D009419:Nerve Tissue Proteins / Q000502:physiology*; D011485:Protein Binding
D051339:Actin Depolymerizing Factors; D000199:Actins; D003432:Cross-Linking Reagents; D008840:Microfilament Proteins; D009124:Muscle Proteins; D009419:Nerve Tissue Proteins; D009218:Myosins