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. 1988 Apr 1;251(1):121–127. doi: 10.1042/bj2510121

Studies on the interaction between actin and cofilin purified by a new method.

N Yonezawa 1, E Nishida 1, S Maekawa 1, H Sakai 1
PMCID: PMC1148972  PMID: 3390149

Abstract

Cofilin is a 21,000-Mr actin-binding protein that widely exists in mammalian tissues. (1) A new purification procedure for porcine brain cofilin has been developed that involves (NH4)2SO4 fractionation and sequential chromatographies on Toyo Pearl and butyl-Toyo Pearl hydrophobic columns, hydroxyapatite, phosphocellulose and Sephadex G-75 gel-filtration columns. The purified cofilin bound to F-actin and increased the amount of G-actin to a limited extent, as previously reported [Nishida, Maekawa & Sakai (1984) Biochemistry 23, 5307-5313]. (2) The binding of cofilin to F-actin was scarcely affected by Mg2+, Ca2+ or by calmodulin. However, the binding was diminished by increasing concentrations of KCl, but was only slightly affected by temperature. (3) Cofilin and either alpha-actinin or filamin could bind to F-actin simultaneously with some competition, but the binding of caldesmon to F-actin was markedly inhibited by cofilin. Phalloidin inhibited the binding of cofilin to F-actin, and protected F-actin from depolymerization by cofilin.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barden J. A., Miki M., Hambly B. D., Dos Remedios C. G. Localization of the phalloidin and nucleotide-binding sites on actin. Eur J Biochem. 1987 Feb 2;162(3):583–588. doi: 10.1111/j.1432-1033.1987.tb10679.x. [DOI] [PubMed] [Google Scholar]
  2. Bretscher A. Smooth muscle caldesmon. Rapid purification and F-actin cross-linking properties. J Biol Chem. 1984 Oct 25;259(20):12873–12880. [PubMed] [Google Scholar]
  3. Burridge K., Feramisco J. R. Non-muscle alpha actinins are calcium-sensitive actin-binding proteins. Nature. 1981 Dec 10;294(5841):565–567. doi: 10.1038/294565a0. [DOI] [PubMed] [Google Scholar]
  4. Coluccio L. M., Tilney L. G. Phalloidin enhances actin assembly by preventing monomer dissociation. J Cell Biol. 1984 Aug;99(2):529–535. doi: 10.1083/jcb.99.2.529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dancker P., Löw I., Hasselbach W., Wieland T. Interaction of actin with phalloidin: polymerization and stabilization of F-actin. Biochim Biophys Acta. 1975 Aug 19;400(2):407–414. doi: 10.1016/0005-2795(75)90196-8. [DOI] [PubMed] [Google Scholar]
  6. Feramisco J. R., Burridge K. A rapid purification of alpha-actinin, filamin, and a 130,000-dalton protein from smooth muscle. J Biol Chem. 1980 Feb 10;255(3):1194–1199. [PubMed] [Google Scholar]
  7. Harwell O. D., Sweeney M. L., Kirkpatrick F. H. Conformation changes of actin during formation of filaments and paracrystals and upon interaction with DNase I, cytochalasin B, and phalloidin. J Biol Chem. 1980 Feb 10;255(3):1210–1220. [PubMed] [Google Scholar]
  8. Korn E. D. Actin polymerization and its regulation by proteins from nonmuscle cells. Physiol Rev. 1982 Apr;62(2):672–737. doi: 10.1152/physrev.1982.62.2.672. [DOI] [PubMed] [Google Scholar]
  9. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  10. Lengsfeld A. M., Löw I., Wieland T., Dancker P., Hasselbach W. Interaction of phalloidin with actin. Proc Natl Acad Sci U S A. 1974 Jul;71(7):2803–2807. doi: 10.1073/pnas.71.7.2803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Maekawa S., Nishida E., Ohta Y., Sakai H. Isolation of low molecular weight actin-binding proteins from porcine brain. J Biochem. 1984 Feb;95(2):377–385. doi: 10.1093/oxfordjournals.jbchem.a134618. [DOI] [PubMed] [Google Scholar]
  12. Mimura N., Asano A. Ca2+-sensitive gelation of actin filaments by a new protein factor. Nature. 1979 Nov 1;282(5734):44–48. doi: 10.1038/282044a0. [DOI] [PubMed] [Google Scholar]
  13. Muneyuki E., Nishida E., Sutoh K., Sakai H. Purification of cofilin, a 21,000 molecular weight actin-binding protein, from porcine kidney and identification of the cofilin-binding site in the actin sequence. J Biochem. 1985 Feb;97(2):563–568. doi: 10.1093/oxfordjournals.jbchem.a135091. [DOI] [PubMed] [Google Scholar]
  14. Nishida E., Iida K., Yonezawa N., Koyasu S., Yahara I., Sakai H. Cofilin is a component of intranuclear and cytoplasmic actin rods induced in cultured cells. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5262–5266. doi: 10.1073/pnas.84.15.5262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nishida E., Kumagai H., Ohtsuki I., Sakai H. The interactions between calcium-dependent regulator protein of cyclic nucleotide phosphodiesterase and microtubule proteins. I. Effect of calcium-dependent regulator protein on the calcium sensitivity of microtubule assembly. J Biochem. 1979 May;85(5):1257–1266. [PubMed] [Google Scholar]
  16. Nishida E., Maekawa S., Sakai H. Cofilin, a protein in porcine brain that binds to actin filaments and inhibits their interactions with myosin and tropomyosin. Biochemistry. 1984 Oct 23;23(22):5307–5313. doi: 10.1021/bi00317a032. [DOI] [PubMed] [Google Scholar]
  17. Nishida E. Opposite effects of cofilin and profilin from porcine brain on rate of exchange of actin-bound adenosine 5'-triphosphate. Biochemistry. 1985 Feb 26;24(5):1160–1164. doi: 10.1021/bi00326a015. [DOI] [PubMed] [Google Scholar]
  18. Pollard T. D., Cooper J. A. Actin and actin-binding proteins. A critical evaluation of mechanisms and functions. Annu Rev Biochem. 1986;55:987–1035. doi: 10.1146/annurev.bi.55.070186.005011. [DOI] [PubMed] [Google Scholar]
  19. Sobue K., Muramoto Y., Fujita M., Kakiuchi S. Purification of a calmodulin-binding protein from chicken gizzard that interacts with F-actin. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5652–5655. doi: 10.1073/pnas.78.9.5652. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Spudich J. A., Watt S. The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin. J Biol Chem. 1971 Aug 10;246(15):4866–4871. [PubMed] [Google Scholar]
  21. Sutoh K., Mabuchi I. Improved method for mapping the binding site of an actin-binding protein in the actin sequence. Use of a site-directed antibody against the N-terminal region of actin as a probe of its N-terminus. Biochemistry. 1986 Oct 7;25(20):6186–6192. doi: 10.1021/bi00368a053. [DOI] [PubMed] [Google Scholar]
  22. Suzuki A., Goll D. E., Singh I., Allen R. E., Robson R. M., Stromer M. H. Some properties of purified skeletal muscle alpha-actinin. J Biol Chem. 1976 Nov 10;251(21):6860–6870. [PubMed] [Google Scholar]
  23. Vandekerckhove J., Deboben A., Nassal M., Wieland T. The phalloidin binding site of F-actin. EMBO J. 1985 Nov;4(11):2815–2818. doi: 10.1002/j.1460-2075.1985.tb04008.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wang K. Filamin, a new high-molecular-weight protein found in smooth muscle and nonmuscle cells. Purification and properties of chicken gizzard filamin. Biochemistry. 1977 May 3;16(9):1857–1865. doi: 10.1021/bi00628a015. [DOI] [PubMed] [Google Scholar]
  25. Wang K., Singer S. J. Interaction of filamin with f-actin in solution. Proc Natl Acad Sci U S A. 1977 May;74(5):2021–2025. doi: 10.1073/pnas.74.5.2021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Weeds A. Actin-binding proteins--regulators of cell architecture and motility. Nature. 1982 Apr 29;296(5860):811–816. doi: 10.1038/296811a0. [DOI] [PubMed] [Google Scholar]
  27. Wieland T., de Vries J. X., Schäfer A., Faulstich H. Spectroscopic evidence for the interaction of phalloidin with actin. FEBS Lett. 1975 Jun 1;54(1):73–75. doi: 10.1016/0014-5793(75)81071-4. [DOI] [PubMed] [Google Scholar]
  28. Yin H. L., Zaner K. S., Stossel T. P. Ca2+ control of actin gelation. Interaction of gelsolin with actin filaments and regulation of actin gelation. J Biol Chem. 1980 Oct 10;255(19):9494–9500. [PubMed] [Google Scholar]
  29. Yonezawa N., Nishida E., Koyasu S., Maekawa S., Ohta Y., Yahara I., Sakai H. Distribution among tissues and intracellular localization of cofilin, a 21kDa actin-binding protein. Cell Struct Funct. 1987 Oct;12(5):443–452. doi: 10.1247/csf.12.443. [DOI] [PubMed] [Google Scholar]
  30. Yonezawa N., Nishida E., Sakai H. pH control of actin polymerization by cofilin. J Biol Chem. 1985 Nov 25;260(27):14410–14412. [PubMed] [Google Scholar]

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