Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1976 Jul 1;70(1):123–143. doi: 10.1083/jcb.70.1.123

The contractile basis of ameboid movement. II. Structure and contractility of motile extracts and plasmalemma-ectoplasm ghosts

PMCID: PMC2109811  PMID: 6480

Abstract

The role of calcium and magnesium-ATP on the structure and contractility in motile extracts of Amoeba proteus and plasmalemma- ectoplasm "ghosts" of Chaos carolinensis has been investigated by correlating light and electron microscope observations with turbidity and birefringence measurements. The extract is nonmotile and contains very few F-actin filaments and myosin aggregates when prepared in the presence of both low calcium ion and ATP concentrations at an ionic strength of I = 0.05, pH 6.8. The addition of 1.0 mM magnesium chloride, 1.0 mM ATP, in the presence of a low calcium ion concentration (relaxation solution) induced the formation of some fibrous bundles of actin without contracting, whereas the addition of a micromolar concentration of calcium in addition to 1.0 mM magnesium-ATP (contraction solution) (Taylor, D. L., J. S. Condeelis, P. L. Moore, and R. D. Allen. 1973. J. Cell Biol. 59:378-394) initiated the formation of large arrays of F-actin filaments followed by contractions. Furthermore, plasmalemma-ectoplasm ghosts prepared in the relaxation solution exhibited very few straight F-actin filaments and myosin aggregates. In contrast, plasmalemmaectoplasm ghosts treated with the contraction solution contained many straight F-actin filaments and myosin aggregates. The increase in the structure of ameba cytoplasm at the endoplasm-ectoplasm interface can be explained by a combination of the transformation of actin from a less filamentous to a more structured filamentous state possibly involving the cross-linking of actin to form fibrillar arrays (see above-mentioned reference) followed by contractions of the actin and myosin along an undetermined distance of the endoplasm and/or ectoplasm.

Full Text

The Full Text of this article is available as a PDF (7.2 MB).

Selected References

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

  1. ALLEN R. D., COOLEDGE J. W., HALL P. J. Streaming in cytoplasm dissociated from the giant amoeba, Chaos chaos. Nature. 1960 Sep 10;187:896–899. doi: 10.1038/187896a0. [DOI] [PubMed] [Google Scholar]
  2. Behnke O., Kristensen B. I., Nielsen L. E. Electron microscopical observations on actinoid and myosinoid filaments in blood platelets. J Ultrastruct Res. 1971 Nov;37(3):351–369. doi: 10.1016/s0022-5320(71)80129-6. [DOI] [PubMed] [Google Scholar]
  3. Comly L. T. Microfilaments in Chaos carolinensis. Membrane association, distribution, and heavy meromyosin binding in the glycerinated cell. J Cell Biol. 1973 Jul;58(1):230–237. doi: 10.1083/jcb.58.1.230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Eckert B. S., McGee-Russell S. M. The patterned organization of thick and thin microfilaments in the contracting pseudopod of Difflugia. J Cell Sci. 1973 Nov;13(3):727–739. doi: 10.1242/jcs.13.3.727. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. GRIFFIN J. L. An improved mass culture method for the large, free-living amebae. Exp Cell Res. 1960 Oct;21:170–178. doi: 10.1016/0014-4827(60)90358-x. [DOI] [PubMed] [Google Scholar]
  7. Gaskin F., Cantor C. R., Shelanski M. L. Turbidimetric studies of the in vitro assembly and disassembly of porcine neurotubules. J Mol Biol. 1974 Nov 15;89(4):737–755. doi: 10.1016/0022-2836(74)90048-5. [DOI] [PubMed] [Google Scholar]
  8. Harris J. R. Further studies on the proteins released from haemoglobin-free erythrocyte ghosts at low ionic strength. Biochim Biophys Acta. 1971 Mar 23;229(3):761–770. doi: 10.1016/0005-2795(71)90294-7. [DOI] [PubMed] [Google Scholar]
  9. Harris J. R., Maddy A. H. An electron microscopic study of some protein fractions from bovine erythrocyte ghosts. J Ultrastruct Res. 1974 Aug;48(2):190–200. doi: 10.1016/s0022-5320(74)80076-6. [DOI] [PubMed] [Google Scholar]
  10. Harris J. R. Some negative contrast staining features of a protein from erythrocyte ghosts. J Mol Biol. 1969 Dec 14;46(2):329–335. doi: 10.1016/0022-2836(69)90425-2. [DOI] [PubMed] [Google Scholar]
  11. Hinssen H., D'Haese J. Filament formation by slime mould myosin isolated at low ionic strength. J Cell Sci. 1974 Jun;15(1):113–129. doi: 10.1242/jcs.15.1.113. [DOI] [PubMed] [Google Scholar]
  12. Holberton D. V., Preston T. M. Arrays of thick filaments in ATP-activated Amoeba model cells. Exp Cell Res. 1970 Oct;62(2):473–477. doi: 10.1016/0014-4827(70)90581-1. [DOI] [PubMed] [Google Scholar]
  13. Kane R. E. Preparation and purification of polymerized actin from sea urchin egg extracts. J Cell Biol. 1975 Aug;66(2):305–315. doi: 10.1083/jcb.66.2.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. LOWEY S., COHEN C. Studies on the structure of myosin. J Mol Biol. 1962 Apr;4:293–308. doi: 10.1016/s0022-2836(62)80007-2. [DOI] [PubMed] [Google Scholar]
  15. Moore P. L., Condeelis J. S., Taylor D. L., Allen R. D. A method for the morphological identification of contractile filaments in single cells. Exp Cell Res. 1973 Aug;80(2):493–495. doi: 10.1016/0014-4827(73)90332-7. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Pollard T. D., Ito S. Cytoplasmic filaments of Amoeba proteus. I. The role of filaments in consistency changes and movement. J Cell Biol. 1970 Aug;46(2):267–289. doi: 10.1083/jcb.46.2.267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pollard T. D., Korn E. D. Filaments of Amoeba proteus. II. Binding of heavy meromyosin by thin filaments in motile cytoplasmic extracts. J Cell Biol. 1971 Jan;48(1):216–219. doi: 10.1083/jcb.48.1.216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. SIMARD-DUQUESNE N., COUILLARD P. Ameboid movement, I. Reactivation of glycerinated models of Amoeba proteus with adenosinetriphosphate. Exp Cell Res. 1962 Oct;28:85–91. doi: 10.1016/0014-4827(62)90314-2. [DOI] [PubMed] [Google Scholar]
  20. Schroeder T. E. Actin in dividing cells: contractile ring filaments bind heavy meromyosin. Proc Natl Acad Sci U S A. 1973 Jun;70(6):1688–1692. doi: 10.1073/pnas.70.6.1688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Stossel T. P., Hartwig J. H. Interactions between actin, myosin, and an actin-binding protein from rabbit alveolar macrophages. Alveolar macrophage myosin Mg-2+-adenosine triphosphatase requires a cofactor for activation by actin. J Biol Chem. 1975 Jul 25;250(14):5706–5712. [PubMed] [Google Scholar]
  22. THOMPSON C. M., WOLPERT L. THE ISOLATION OF MOTILE CYTOPLASM FROM AMOEBA PROTEUS. Exp Cell Res. 1963 Oct;32:156–160. doi: 10.1016/0014-4827(63)90078-8. [DOI] [PubMed] [Google Scholar]
  23. Taylor D. L., Condeelis J. S., Moore P. L., Allen R. D. The contractile basis of amoeboid movement. I. The chemical control of motility in isolated cytoplasm. J Cell Biol. 1973 Nov;59(2 Pt 1):378–394. doi: 10.1083/jcb.59.2.378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tilney L. G. Actin filaments in the acrosomal reaction of Limulus sperm. Motion generated by alterations in the packing of the filaments. J Cell Biol. 1975 Feb;64(2):289–310. doi: 10.1083/jcb.64.2.289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tilney L. G., Detmers P. Actin in erythrocyte ghosts and its association with spectrin. Evidence for a nonfilamentous form of these two molecules in situ. J Cell Biol. 1975 Sep;66(3):508–520. doi: 10.1083/jcb.66.3.508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tilney L. G., Hatano S., Ishikawa H., Mooseker M. S. The polymerization of actin: its role in the generation of the acrosomal process of certain echinoderm sperm. J Cell Biol. 1973 Oct;59(1):109–126. doi: 10.1083/jcb.59.1.109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Tilney L. G., Mooseker M. Actin in the brush-border of epithelial cells of the chicken intestine. Proc Natl Acad Sci U S A. 1971 Oct;68(10):2611–2615. doi: 10.1073/pnas.68.10.2611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  29. Wohlman A., Allen R. D. Structural organization associated with pseudopod extension and contraction during cell locomotion in Difflugia. J Cell Sci. 1968 Mar;3(1):105–114. doi: 10.1242/jcs.3.1.105. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES