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. 1984 May 1;98(5):1842–1850. doi: 10.1083/jcb.98.5.1842

Monoclonal antibodies to dynein subunits reveal the existence of cytoplasmic antigens in sea urchin egg

PMCID: PMC2113199  PMID: 6233290

Abstract

Monoclonal antibodies directed against subunits of a sea urchin flagellar dynein were used to test for the presence of cytoplasmic antigens in preparations of fertilized eggs and mitotic apparati . A 9- 10 S complex composed of 330,000-, 134,000-, and 126,000-mol-wt subunits was isolated from outer arms of Strongylocentrotus purpuratus sperm flagella and used to characterize the antibodies. Seven monospecific antibodies to the 330,000 subunit and two against the 134,000 subunit of the 9-10 S complex were identified by binding to nitrocellulose blots of electrophoretograms resolving polypeptides from different dynein preparations. The antibodies were applied also to blots of polypeptides from fertilized sea urchin egg at the first metaphase and a cellular fraction of mitotic apparati . Three of the antibodies to the 330,000 subunit bound to a cytoplasmic polypeptide of approximately the same molecular weight and the two antibodies to the smaller subunits recognized a polypeptide of 124,000 apparent molecular weight. Both antigens appeared to be enriched in the fraction containing mitotic apparati . These results indicate that polypeptides similar to two subunits of the 9-10 S complex are present in eggs at metaphase, and they are apparently associated with the mitotic apparatus.

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Selected References

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

  1. Bell C. W., Fronk E., Gibbons I. R. Polypeptide subunits of dynein 1 from sea urchin sperm flagella. J Supramol Struct. 1979;11(3):311–317. doi: 10.1002/jss.400110305. [DOI] [PubMed] [Google Scholar]
  2. Cande W. Z., Wolniak S. M. Chromosome movement in lysed mitotic cells is inhibited by vanadate. J Cell Biol. 1978 Nov;79(2 Pt 1):573–580. doi: 10.1083/jcb.79.2.573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Gibbons B. H., Gibbons I. R. Relationship between the latent adenosine triphosphatase state of dynein 1 and its ability to recombine functionally with KCl-extracted sea urchin sperm flagella. J Biol Chem. 1979 Jan 10;254(1):197–201. [PubMed] [Google Scholar]
  4. Gibbons I. R., Fronk E. A latent adenosine triphosphatase form of dynein 1 from sea urchin sperm flagella. J Biol Chem. 1979 Jan 10;254(1):187–196. [PubMed] [Google Scholar]
  5. HUNTER W. M., GREENWOOD F. C. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. doi: 10.1038/194495a0. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Hisanaga S., Sakai H. Cytoplasmic dynein of the sea urchin egg. II. Purification, characterization and interactions with microtubules and Ca-calmodulin. J Biochem. 1983 Jan;93(1):87–98. doi: 10.1093/oxfordjournals.jbchem.a134182. [DOI] [PubMed] [Google Scholar]
  8. Huang B., Piperno G., Luck D. J. Paralyzed flagella mutants of Chlamydomonas reinhardtii. Defective for axonemal doublet microtubule arms. J Biol Chem. 1979 Apr 25;254(8):3091–3099. [PubMed] [Google Scholar]
  9. Klee C. B., Crouch T. H., Richman P. G. Calmodulin. Annu Rev Biochem. 1980;49:489–515. doi: 10.1146/annurev.bi.49.070180.002421. [DOI] [PubMed] [Google Scholar]
  10. Mabuchi I. ATPase in the cortical layer of sea urchin egg: its properties and interaction with cortex protein. Biochim Biophys Acta. 1973 Feb 28;297(2):317–332. doi: 10.1016/0304-4165(73)90079-2. [DOI] [PubMed] [Google Scholar]
  11. McIntosh J. R., Cande W. Z., Snyder J. A. Structure and physiology of the mammalian mitotic spindle. Soc Gen Physiol Ser. 1975;30:31–76. [PubMed] [Google Scholar]
  12. Piperno G., Luck D. J. Axonemal adenosine triphosphatases from flagella of Chlamydomonas reinhardtii. Purification of two dyneins. J Biol Chem. 1979 Apr 25;254(8):3084–3090. [PubMed] [Google Scholar]
  13. Piperno G., Luck D. J. Inner arm dyneins from flagella of Chlamydomonas reinhardtii. Cell. 1981 Dec;27(2 Pt 1):331–340. doi: 10.1016/0092-8674(81)90416-5. [DOI] [PubMed] [Google Scholar]
  14. Piperno G., Luck D. J. Microtubular proteins of Chlamydomonas reinhardtii. An immunochemical study based on the use of an antibody specific for the beta-tubulin subunit. J Biol Chem. 1977 Jan 10;252(1):383–391. [PubMed] [Google Scholar]
  15. Pratt M. M., Otter T., Salmon E. D. Dynein-like Mg2+-ATPase in mitotic spindles isolated from sea urchin embryos (Strongylocentrotus droebachiensis). J Cell Biol. 1980 Sep;86(3):738–745. doi: 10.1083/jcb.86.3.738. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pratt M. M. The identification of a dynein ATPase in unfertilized sea urchin eggs. Dev Biol. 1980 Feb;74(2):364–378. doi: 10.1016/0012-1606(80)90438-8. [DOI] [PubMed] [Google Scholar]
  17. Salmon E. D., Segall R. R. Calcium-labile mitotic spindles isolated from sea urchin eggs (Lytechinus variegatus). J Cell Biol. 1980 Aug;86(2):355–365. doi: 10.1083/jcb.86.2.355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Tang W. J., Bell C. W., Sale W. S., Gibbons I. R. Structure of the dynein-1 outer arm in sea urchin sperm flagella. I. Analysis by separation of subunits. J Biol Chem. 1982 Jan 10;257(1):508–515. [PubMed] [Google Scholar]
  20. Telzer B. R., Haimo L. T. Decoration of spindle microtubules with Dynein: evidence for uniform polarity. J Cell Biol. 1981 May;89(2):373–378. doi: 10.1083/jcb.89.2.373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. de StGroth S. F., Scheidegger D. Production of monoclonal antibodies: strategy and tactics. J Immunol Methods. 1980;35(1-2):1–21. doi: 10.1016/0022-1759(80)90146-5. [DOI] [PubMed] [Google Scholar]

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