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. 1984 Jul 1;99(1 Pt 1):71–78. doi: 10.1083/jcb.99.1.71

A membrane cytoskeleton from Dictyostelium discoideum. III. Plasma membrane fragments bind predominantly to the sides of actin filaments

PMCID: PMC2275626  PMID: 6610683

Abstract

The binding between sonicated Dictyostelium discoideum plasma membrane fragments and F-actin on Sephacryl S-1000 beads was found to be competitively inhibited by myosin subfragment-1. This inhibition is MgATP-sensitive, exhibits a Ki of approximately 5 X 10(-8) M, and is reciprocal, since membranes inhibit the binding of 125I-heavy meromyosin to F-actin on beads. These experiments demonstrate that membrane binding and S-1 binding to F-actin on beads are mutually exclusive and, therefore, that the membrane fragments bind predominantly to the sides, rather than to the ends, of the actin filaments. This conclusion is supported by electron micrographs that show many lateral associations between membrane fragments and bead- associated actin filaments. Such lateral associations could play an important role in the organization and lateral movement of membrane proteins by the cytomusculature.

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Selected References

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  1. Begg D. A., Rodewald R., Rebhun L. I. The visualization of actin filament polarity in thin sections. Evidence for the uniform polarity of membrane-associated filaments. J Cell Biol. 1978 Dec;79(3):846–852. doi: 10.1083/jcb.79.3.846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bolton A. E., Hunter W. M. The labelling of proteins to high specific radioactivities by conjugation to a 125I-containing acylating agent. Biochem J. 1973 Jul;133(3):529–539. doi: 10.1042/bj1330529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bourguignon L. Y., Singer S. J. Transmembrane interactions and the mechanism of capping of surface receptors by their specific ligands. Proc Natl Acad Sci U S A. 1977 Nov;74(11):5031–5035. doi: 10.1073/pnas.74.11.5031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Braun J., Fujiwara K., Pollard T. D., Unanue E. R. Two distinct mechanisms for redistribution of lymphocyte surface macromolecules. I. Relationship to cytoplasmic myosin. J Cell Biol. 1978 Nov;79(2 Pt 1):409–418. doi: 10.1083/jcb.79.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brenner S. L., Korn E. D. Spectrin-actin interaction. Phosphorylated and dephosphorylated spectrin tetramer cross-link F-actin. J Biol Chem. 1979 Sep 10;254(17):8620–8627. [PubMed] [Google Scholar]
  6. Cohen C. M., Tyler J. M., Branton D. Spectrin-actin associations studied by electron microscopy of shadowed preparations. Cell. 1980 Oct;21(3):875–883. doi: 10.1016/0092-8674(80)90451-1. [DOI] [PubMed] [Google Scholar]
  7. Condeelis J. Isolation of concanavalin A caps during various stages of formation and their association with actin and myosin. J Cell Biol. 1979 Mar;80(3):751–758. doi: 10.1083/jcb.80.3.751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. DIXON M. The determination of enzyme inhibitor constants. Biochem J. 1953 Aug;55(1):170–171. doi: 10.1042/bj0550170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Edds K. T. Microfilament bundles. I. Formation with uniform polarity. Exp Cell Res. 1977 Sep;108(2):452–456. doi: 10.1016/s0014-4827(77)80056-6. [DOI] [PubMed] [Google Scholar]
  10. Edelman G. M. Surface modulation in cell recognition and cell growth. Science. 1976 Apr 16;192(4236):218–226. doi: 10.1126/science.769162. [DOI] [PubMed] [Google Scholar]
  11. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  12. Flanagan J., Koch G. L. Cross-linked surface Ig attaches to actin. Nature. 1978 May 25;273(5660):278–281. doi: 10.1038/273278a0. [DOI] [PubMed] [Google Scholar]
  13. Fowler V. M., Luna E. J., Hargreaves W. R., Taylor D. L., Branton D. Spectrin promotes the association of F-actin with the cytoplasmic surface of the human erythrocyte membrane. J Cell Biol. 1981 Feb;88(2):388–395. doi: 10.1083/jcb.88.2.388. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fowler V. M., Pollard H. B. Chromaffin granule membrane-F-actin interactions are calcium sensitive. Nature. 1982 Jan 28;295(5847):336–339. doi: 10.1038/295336a0. [DOI] [PubMed] [Google Scholar]
  15. Fowler V., Taylor D. L. Spectrin plus band 4.1 cross-link actin. Regulation by micromolar calcium. J Cell Biol. 1980 May;85(2):361–376. doi: 10.1083/jcb.85.2.361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fraser A. B., Eisenberg E., Kielley W. W., Carlson F. D. The interaction of heavy meromyosin and subfragment 1 with actin. Physical measurements in the presence and absence of adenosine triphosphate. Biochemistry. 1975 May 20;14(10):2207–2214. doi: 10.1021/bi00681a025. [DOI] [PubMed] [Google Scholar]
  17. Greene L. E., Eisenberg E. Dissociation of the actin.subfragment 1 complex by adenyl-5'-yl imidodiphosphate, ADP, and PPi. J Biol Chem. 1980 Jan 25;255(2):543–548. [PubMed] [Google Scholar]
  18. Highsmith S., Mendelson R. A., Morales M. F. Affinity of myosin S-1 for F-actin, measured by time-resolved fluorescence anisotropy. Proc Natl Acad Sci U S A. 1976 Jan;73(1):133–137. doi: 10.1073/pnas.73.1.133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hirokawa N., Tilney L. G. Interactions between actin filaments and between actin filaments and membranes in quick-frozen and deeply etched hair cells of the chick ear. J Cell Biol. 1982 Oct;95(1):249–261. doi: 10.1083/jcb.95.1.249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Howe C. L., Mooseker M. S. Characterization of the 110-kdalton actin-calmodulin-, and membrane-binding protein from microvilli of intestinal epithelial cells. J Cell Biol. 1983 Oct;97(4):974–985. doi: 10.1083/jcb.97.4.974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Isenberg G., Leonard K., Jockusch B. M. Structural aspects of vinculin-actin interactions. J Mol Biol. 1982 Jun 25;158(2):231–249. doi: 10.1016/0022-2836(82)90431-4. [DOI] [PubMed] [Google Scholar]
  22. Ishikawa H., Bischoff R., Holtzer H. Formation of arrowhead complexes with heavy meromyosin in a variety of cell types. J Cell Biol. 1969 Nov;43(2):312–328. [PMC free article] [PubMed] [Google Scholar]
  23. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  24. 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]
  25. Luna E. J., Fowler V. M., Swanson J., Branton D., Taylor D. L. A membrane cytoskeleton from Dictyostelium discoideum. I. Identification and partial characterization of an actin-binding activity. J Cell Biol. 1981 Feb;88(2):396–409. doi: 10.1083/jcb.88.2.396. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Luna E. J., Goodloe-Holland C. M., Ingalls H. M. A membrane cytoskeleton from Dictyostelium discoideum. II. Integral proteins mediate the binding of plasma membranes to F-actin affinity beads. J Cell Biol. 1984 Jul;99(1 Pt 1):58–70. doi: 10.1083/jcb.99.1.58. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Luna E. J., Wang Y. L., Voss E. W., Jr, Branton D., Taylor D. L. A stable, high capacity, F-actin affinity column. J Biol Chem. 1982 Nov 10;257(21):13095–13100. [PubMed] [Google Scholar]
  28. Margossian S. S., Lowey S. Interaction of myosin subfragments with F-actin. Biochemistry. 1978 Dec 12;17(25):5431–5439. doi: 10.1021/bi00618a017. [DOI] [PubMed] [Google Scholar]
  29. Marston S., Weber A. The dissociation constant of the actin-heavy meromyosin subfragment-1 complex. Biochemistry. 1975 Aug 26;14(17):3868–3873. doi: 10.1021/bi00688a021. [DOI] [PubMed] [Google Scholar]
  30. McNutt N. S. A thin-section and freeze-fracture study of microfilament-membrane attachments in choroid plexus and intestinal microvilli. J Cell Biol. 1978 Dec;79(3):774–787. doi: 10.1083/jcb.79.3.774. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Mooseker M. S., Tilney L. G. Organization of an actin filament-membrane complex. Filament polarity and membrane attachment in the microvilli of intestinal epithelial cells. J Cell Biol. 1975 Dec;67(3):725–743. doi: 10.1083/jcb.67.3.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Mornet D., Bertrand R., Pantel P., Audemard E., Kassab R. Structure of the actin-myosin interface. Nature. 1981 Jul 23;292(5821):301–306. doi: 10.1038/292301a0. [DOI] [PubMed] [Google Scholar]
  33. Small J. V., Isenberg G., Celis J. E. Polarity of actin at the leading edge of cultured cells. Nature. 1978 Apr 13;272(5654):638–639. doi: 10.1038/272638a0. [DOI] [PubMed] [Google Scholar]
  34. Stein L. A., Schwarz R. P., Jr, Chock P. B., Eisenberg E. Mechanism of actomyosin adenosine triphosphatase. Evidence that adenosine 5'-triphosphate hydrolysis can occur without dissociation of the actomyosin complex. Biochemistry. 1979 Sep 4;18(18):3895–3909. doi: 10.1021/bi00585a009. [DOI] [PubMed] [Google Scholar]
  35. Tsukita S., Tsukita S., Ishikawa H., Sato S., Nakao M. Electron microscopic study of reassociation of spectrin and actin with the human erythrocyte membrane. J Cell Biol. 1981 Jul;90(1):70–77. doi: 10.1083/jcb.90.1.70. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Weeds A. G., Pope B. Studies on the chymotryptic digestion of myosin. Effects of divalent cations on proteolytic susceptibility. J Mol Biol. 1977 Apr;111(2):129–157. doi: 10.1016/s0022-2836(77)80119-8. [DOI] [PubMed] [Google Scholar]
  37. White H. D., Taylor E. W. Energetics and mechanism of actomyosin adenosine triphosphatase. Biochemistry. 1976 Dec 28;15(26):5818–5826. doi: 10.1021/bi00671a020. [DOI] [PubMed] [Google Scholar]
  38. YOUNG D. M., HIMMELFARB S., HARRINGTON W. F. ON THE STRUCTURAL ASSEMBLY OF THE POLYPEPTIDE CHAINS OF HEAVY MEROMYOSIN. J Biol Chem. 1965 Jun;240:2428–2436. [PubMed] [Google Scholar]

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