Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1984 Oct 1;99(4):1527–1533. doi: 10.1083/jcb.99.4.1527

ATP-induced gelation--contraction of microtubules assembled in vitro

PMCID: PMC2113323  PMID: 6480701

Abstract

We report here an ATP-dependent formation and contraction, or syneresis, of a microtubule gel using microtubule proteins prepared from calf brains. Gel contraction is typically observable 15-30 min after ATP addition to microtubules assembled to steady state, and is complete after approximately 60 min, at which time the gel volume is reduced by as much as 75%. In contracted gels, microtubule bundles and aster-like structures are observable. Gelation-contraction requires only microtubule proteins present after purification by three cycles of assembly and disassembly.

Full Text

The Full Text of this article is available as a PDF (1.1 MB).

Selected References

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

  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  2. Cande W. Z. Nucleotide requirements for anaphase chromosome movements in permeabilized mitotic cells: anaphase B but not anaphase A requires ATP. Cell. 1982 Jan;28(1):15–22. doi: 10.1016/0092-8674(82)90370-1. [DOI] [PubMed] [Google Scholar]
  3. Caplow M., Zeeberg B. Dynamic properties of microtubules at steady state in the presence of taxol. Eur J Biochem. 1982 Oct;127(2):319–324. doi: 10.1111/j.1432-1033.1982.tb06873.x. [DOI] [PubMed] [Google Scholar]
  4. Ellisman M. H., Porter K. R. Microtrabecular structure of the axoplasmic matrix: visualization of cross-linking structures and their distribution. J Cell Biol. 1980 Nov;87(2 Pt 1):464–479. doi: 10.1083/jcb.87.2.464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gaskin F., Kramer S. B., Cantor C. R., Adelstein R., Shelanski M. L. A dynein-like protein associated with neurotubules. FEBS Lett. 1974 Apr 1;40(2):281–286. doi: 10.1016/0014-5793(74)80244-9. [DOI] [PubMed] [Google Scholar]
  6. Griffith L. M., Pollard T. D. Evidence for actin filament-microtubule interaction mediated by microtubule-associated proteins. J Cell Biol. 1978 Sep;78(3):958–965. doi: 10.1083/jcb.78.3.958. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Haimo L. T., Telzer B. R., Rosenbaum J. L. Dynein binds to and crossbridges cytoplasmic microtubules. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5759–5763. doi: 10.1073/pnas.76.11.5759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hirokawa N. Cross-linker system between neurofilaments, microtubules, and membranous organelles in frog axons revealed by the quick-freeze, deep-etching method. J Cell Biol. 1982 Jul;94(1):129–142. doi: 10.1083/jcb.94.1.129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ishikawa M., Murofushi H., Sakai H. Bundling of microtubules in vitro by fodrin. J Biochem. 1983 Oct;94(4):1209–1217. doi: 10.1093/oxfordjournals.jbchem.a134466. [DOI] [PubMed] [Google Scholar]
  10. Jameson L., Frey T., Zeeberg B., Dalldorf F., Caplow M. Inhibition of microtubule assembly by phosphorylation of microtubule-associated proteins. Biochemistry. 1980 May 27;19(11):2472–2479. doi: 10.1021/bi00552a027. [DOI] [PubMed] [Google Scholar]
  11. Kirschner M. W. Implications of treadmilling for the stability and polarity of actin and tubulin polymers in vivo. J Cell Biol. 1980 Jul;86(1):330–334. doi: 10.1083/jcb.86.1.330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kumagai H., Sakai H. A porcine brain protein (35 K protein) which bundles microtubules and its identification as glyceraldehyde 3-phosphate dehydrogenase. J Biochem. 1983 May;93(5):1259–1269. doi: 10.1093/oxfordjournals.jbchem.a134260. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Margolis R. L., Wilson L. Opposite end assembly and disassembly of microtubules at steady state in vitro. Cell. 1978 Jan;13(1):1–8. doi: 10.1016/0092-8674(78)90132-0. [DOI] [PubMed] [Google Scholar]
  15. Minami Y., Murofushi H., Sakai H. Interaction of tubulin with neurofilaments: formation of networks by neurofilament-dependent tubulin polymerization. J Biochem. 1982 Sep;92(3):889–898. doi: 10.1093/oxfordjournals.jbchem.a134003. [DOI] [PubMed] [Google Scholar]
  16. Minton A. P. A thermodynamic model for gelation of sickle-cell hemoglobin. J Mol Biol. 1974 Feb 5;82(4):483–498. doi: 10.1016/0022-2836(74)90243-5. [DOI] [PubMed] [Google Scholar]
  17. Murofushi H., Minami Y., Matsumoto G., Sakai H. Bundling of microtubules in vitro by a high molecular weight protein prepared from the squid axon. J Biochem. 1983 Feb;93(2):639–650. doi: 10.1093/oxfordjournals.jbchem.a134220. [DOI] [PubMed] [Google Scholar]
  18. Neal M. W., Florini J. R. A rapid method for desalting small volumes of solution. Anal Biochem. 1973 Sep;55(1):328–330. doi: 10.1016/0003-2697(73)90325-4. [DOI] [PubMed] [Google Scholar]
  19. Runge M. S., Laue T. M., Yphantis D. A., Lifsics M. R., Saito A., Altin M., Reinke K., Williams R. C., Jr ATP-induced formation of an associated complex between microtubules and neurofilaments. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1431–1435. doi: 10.1073/pnas.78.3.1431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Shaw G., Bray D. Movement and extension of isolated growth cones. Exp Cell Res. 1977 Jan;104(1):55–62. doi: 10.1016/0014-4827(77)90068-4. [DOI] [PubMed] [Google Scholar]
  21. Simons T. J. Vanadate--a new tool for biologists. Nature. 1979 Oct 4;281(5730):337–338. doi: 10.1038/281337a0. [DOI] [PubMed] [Google Scholar]
  22. Sloboda R. D., Rosenbaum J. L. Purification and assay of microtubule-associated proteins (MAPs). Methods Enzymol. 1982;85(Pt B):409–416. doi: 10.1016/0076-6879(82)85041-6. [DOI] [PubMed] [Google Scholar]
  23. Sloboda R. D., Rudolph S. A., Rosenbaum J. L., Greengard P. Cyclic AMP-dependent endogenous phosphorylation of a microtubule-associated protein. Proc Natl Acad Sci U S A. 1975 Jan;72(1):177–181. doi: 10.1073/pnas.72.1.177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Summers K. E., Gibbons I. R. Adenosine triphosphate-induced sliding of tubules in trypsin-treated flagella of sea-urchin sperm. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3092–3096. doi: 10.1073/pnas.68.12.3092. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tohyama K., Miller W. G. Network structure in gels of rod-like polypeptides. Nature. 1981 Feb 26;289(5800):813–814. doi: 10.1038/289813a0. [DOI] [PubMed] [Google Scholar]
  26. Tominaga S., Kaziro Y. Adenosine triphosphatases associated with bovine brain microtubules. I. Presence of two distinct ATPases and their partial purification. J Biochem. 1983 Apr;93(4):1085–1092. doi: 10.1093/oxfordjournals.jbchem.a134233. [DOI] [PubMed] [Google Scholar]
  27. Travis J. L., Kenealy J. F., Allen R. D. Studies on the motility of the foraminifera. II. The dynamic microtubular cytoskeleton of the reticulopodial network of Allogromia laticollaris. J Cell Biol. 1983 Dec;97(6):1668–1676. doi: 10.1083/jcb.97.6.1668. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Warren R. H., Brunside B. Microtubules in cone myoid elongation in the teleost retina. J Cell Biol. 1978 Jul;78(1):247–259. doi: 10.1083/jcb.78.1.247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Webb B. C. An ATPase activity associated with brain microtubules. Arch Biochem Biophys. 1979 Nov;198(1):296–303. doi: 10.1016/0003-9861(79)90422-3. [DOI] [PubMed] [Google Scholar]
  30. Zackroff R. V., Weisenberg R. C., Deery W. J. Equilibrium and kinetic analysis of microtubule assembly in the presence of guanosine diphosphate. J Mol Biol. 1980 Jun 5;139(4):641–659. doi: 10.1016/0022-2836(80)90053-4. [DOI] [PubMed] [Google Scholar]

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

RESOURCES