Actin in Triton-treated Cortical Preparations of Unfertilized and Fertilized Sea Urchin Eggs

Overview

Journal J Cell Biol

Specialty Cell Biology

Date 1979 Jul 1

PMID 573270

Citations 17

Authors

A Spudich

J A Spudich

Affiliations

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Abstract

Triton-treated cortical fragments of unfertilized and fertilized sea urchin eggs prepared in the presence of greater than or equal to 5 mM EGTA contain 15-30% of the total egg actin. However, actin filaments are not readily apparent by electron microscopy on the cortical fragments of unfertilized eggs but are numerous on those of fertilized eggs. The majority of the actin associated with cortical fragments of unfertilized eggs is solubilized by dialysis against a low ionic strength buffer at pH 7.5. This soluble actin preparation (less than 50% pure actin) does not form proper filaments in 0.1 M KCl and 3 mM MgCl2, whereas actin purified from this preparation does, as judged by electron microscopy. Optical diffraction analysis reveals that these purified actin filaments have helical parameters very similar to those of muscle actin. Furthermore, the properties of the purified actin with regard to activation of myosin ATPase are similar to those of actin from other cell types. The possibility that actin is maintained in a nonfilamentous form on the inner surface of the unfertilized egg plasma membrane and is induced to assemble upon fertilization is discussed.

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References

AMES G . Resolution of bacterial proteins by polyacrylamide gel electrophoresis on slabs. Membrane, soluble, and periplasmic fractions. J Biol Chem. 1974; 249(2):634-44. View

Harris P . Cortical fibers in fertilized eggs of the sea urchin Strongylocentrotus purpuratus. Exp Cell Res. 1968; 52(2):677-81. DOI: 10.1016/0014-4827(68)90509-0. View

MIKI-NOUMURA T . An actin-like protein of the sea urchin eggs. II. Direct isolation procedure. Dev Growth Differ. 1969; 11(3):219-31. DOI: 10.1111/j.1440-169x.1969.00219.x. View

Spudich J, Clarke M . The contractile proteins of Dictyostelium discoideum. J Supramol Struct. 1974; 2(2-4):150-62. DOI: 10.1002/jss.400020209. View

Spudich J . Biochemical and structural studies of actomyosin-like proteins from non-muscle cells. II. Purification, properties, and membrane association of actin from amoebae of Dictyostelium discoideum. J Biol Chem. 1974; 249(18):6013-20. View

Hartree E . Determination of protein: a modification of the Lowry method that gives a linear photometric response. Anal Biochem. 1972; 48(2):422-7. DOI: 10.1016/0003-2697(72)90094-2. View

Lazarides E, Lindberg U . Actin is the naturally occurring inhibitor of deoxyribonuclease I. Proc Natl Acad Sci U S A. 1974; 71(12):4742-6. PMC: 433972. DOI: 10.1073/pnas.71.12.4742. View

Clarke M, Schatten G, MAZIA D, Spudich J . Visualization of actin fibers associated with the cell membrane in amoebae of Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1975; 72(5):1758-62. PMC: 432625. DOI: 10.1073/pnas.72.5.1758. View

Spudich J, Huxley H, Finch J . Regulation of skeletal muscle contraction. II. Structural studies of the interaction of the tropomyosin-troponin complex with actin. J Mol Biol. 1972; 72(3):619-32. DOI: 10.1016/0022-2836(72)90180-5. View

10.

MIKI-NOUMURA T, Oosawa F . An actin-like protein of the sea urchin eggs. I. Its interaction with myosin from rabbit striated muscle. Exp Cell Res. 1969; 56(2):224-32. DOI: 10.1016/0014-4827(69)90006-8. View

11.

Schroeder T . Actin in dividing cells: contractile ring filaments bind heavy meromyosin. Proc Natl Acad Sci U S A. 1973; 70(6):1688-92. PMC: 433573. DOI: 10.1073/pnas.70.6.1688. View

12.

Johnson J, Epel D . Intracellular pH and activation of sea urchin eggs after fertilisation. Nature. 1976; 262(5570):661-4. DOI: 10.1038/262661a0. View

13.

Ofarrell P . High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975; 250(10):4007-21. PMC: 2874754. View

14.

REBHUN L, Rosenbaum J, Lefebvre P, Smith G . Reversible restoration of the birefringence of cold-treated, isolated mitotic apparatus of surf clam eggs with chick brain tubulin. Nature. 1974; 249(453):113-5. DOI: 10.1038/249113a0. View

15.

Hiramoto Y . Rheological properties of sea urchin eggs. Biorheology. 1970; 6(3):201-34. DOI: 10.3233/bir-1970-6306. View

16.

Runnstrom J . The vitelline membrane and cortical particles in sea urchin eggs and their function in maturation and fertilization. Adv Morphog. 1966; 5:221-325. DOI: 10.1016/b978-1-4831-9952-8.50010-9. View

17.

Kane R . Preparation and purification of polymerized actin from sea urchin egg extracts. J Cell Biol. 1975; 66(2):305-15. PMC: 2109559. DOI: 10.1083/jcb.66.2.305. View

18.

Vacquier V . The isolation of intact cortical granules from sea urchin eggs: calcium lons trigger granule discharge. Dev Biol. 1975; 43(1):62-74. DOI: 10.1016/0012-1606(75)90131-1. View

19.

Epel D, Weaver A, MAZIA D . Methods for revoval of the vitelline membrane of sea urchin eggs. I. Use of dithiothreitol (Cleland Reagent). Exp Cell Res. 1970; 61(1):64-8. DOI: 10.1016/0014-4827(70)90257-0. View

20.

MIKI-NOUMURA T, Kondo H . Polymerization of actin from sea urchin eggs. Exp Cell Res. 1970; 61(1):31-41. DOI: 10.1016/0014-4827(70)90254-5. View