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. 1987 Oct 1;105(4):1649–1662. doi: 10.1083/jcb.105.4.1649

The crystal lattice of Paramecium trichocysts before and after exocytosis by X-ray diffraction and freeze-fracture electron microscopy

PMCID: PMC2114671  PMID: 3667694

Abstract

Paramecium trichocysts are unusual secretory organelles in that: (a) their crystalline contents are built up from a family of low molecular mass acidic proteins; (b) they have a precise, genetically determined shape; and (c) the crystalline trichocyst contents expand rapidly upon exocytosis to give a second, extracellular form which is also an ordered array. We report here the first step of our study of trichocyst structure. We have used a combination of x-ray powder diffraction, freeze-etching, and freeze-fracture electron microscopy of isolated, untreated trichocysts, and density measurements to show that trichocyst contents are indeed protein crystals and to determine the elementary unit cell of both the compact intracellular and the extended extracellular form.

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Selected References

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  1. Adoutte A., Ramanathan R., Lewis R. M., Dute R. R., Ling K. Y., Kung C., Nelson D. L. Biochemical studies of the excitable membrane of Paramecium tetraurelia. III. Proteins of cilia and ciliary membranes. J Cell Biol. 1980 Mar;84(3):717–738. doi: 10.1083/jcb.84.3.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Adoutte A., de Loubresse N. G., Beisson J. Proteolytic cleavage and maturation of the crystalline secretion products of Paramecium. J Mol Biol. 1984 Dec 25;180(4):1065–1081. doi: 10.1016/0022-2836(84)90271-7. [DOI] [PubMed] [Google Scholar]
  3. Aggerbeck L. P., Gulik-Krzywicki T. Studies of lipoproteins by freeze-fracture and etching electron microscopy. Methods Enzymol. 1986;128:457–472. doi: 10.1016/0076-6879(86)28087-8. [DOI] [PubMed] [Google Scholar]
  4. Anderer R., Hausmann K. Properties and structure of isolated extrusive organelles. J Ultrastruct Res. 1977 Jul;60(1):21–26. doi: 10.1016/s0022-5320(77)80037-3. [DOI] [PubMed] [Google Scholar]
  5. Bannister L. H. The structure of trichocysts in Paramecium caudatum. J Cell Sci. 1972 Nov;11(3):899–929. doi: 10.1242/jcs.11.3.899. [DOI] [PubMed] [Google Scholar]
  6. Bilinski M., Plattner H., Matt H. Secretory protein decondensation as a distinct, Ca2+-mediated event during the final steps of exocytosis in Paramecium cells. J Cell Biol. 1981 Jan;88(1):179–188. doi: 10.1083/jcb.88.1.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Caspar D. L., Cohen C., Longley W. Tropomyosin: crystal structure, polymorphism and molecular interactions. J Mol Biol. 1969 Apr 14;41(1):87–107. doi: 10.1016/0022-2836(69)90128-4. [DOI] [PubMed] [Google Scholar]
  8. Cohen C., Caspar D. L., Parry D. A., Lucas R. M. Tropomyosin crystal dynamics. Cold Spring Harb Symp Quant Biol. 1972;36:205–216. doi: 10.1101/sqb.1972.036.01.028. [DOI] [PubMed] [Google Scholar]
  9. Cohen C., Tooney N. M. Crystallisation of a modified fibrinogen. Nature. 1974 Oct 18;251(5476):659–660. doi: 10.1038/251659a0. [DOI] [PubMed] [Google Scholar]
  10. Cohen J., Beisson J. Genetic analysis of the relationships between the cell surface and the nuclei in Paramecium tetraurella. Genetics. 1980 Aug;95(4):797–818. doi: 10.1093/genetics/95.4.797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Garofalo R. S., Gilligan D. M., Satir B. H. Calmodulin antagonists inhibit secretion in Paramecium. J Cell Biol. 1983 Apr;96(4):1072–1081. doi: 10.1083/jcb.96.4.1072. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Garofalo R. S., Satir B. H. Paramecium secretory granule content: quantitative studies on in vitro expansion and its regulation by calcium and pH. J Cell Biol. 1984 Dec;99(6):2193–2199. doi: 10.1083/jcb.99.6.2193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Guraya S. S. Recent progress in the structure, origin, composition, and function of cortical granules in animal egg. Int Rev Cytol. 1982;78:257–360. doi: 10.1016/s0074-7696(08)60108-4. [DOI] [PubMed] [Google Scholar]
  14. Hausmann K. Extrusive organelles in protists. Int Rev Cytol. 1978;52:197–276. doi: 10.1016/s0074-7696(08)60757-3. [DOI] [PubMed] [Google Scholar]
  15. Holmes K. C., Monro R. E. Studies on the structure of parasporal inclusions from Bacillus thuringiensis. J Mol Biol. 1965 Dec;14(2):572–581. doi: 10.1016/s0022-2836(65)80205-4. [DOI] [PubMed] [Google Scholar]
  16. Mathews F. S., Lederer F. Crystallographic study of bakers' yeast cytochrome b2. J Mol Biol. 1976 Apr 25;102(4):853–857. doi: 10.1016/0022-2836(76)90295-3. [DOI] [PubMed] [Google Scholar]
  17. Matt H., Plattner H., Reichel K., Lefort-Tran M., Beisson J. Genetic dissection of the final exocytosis steps in Paramecium tetraurelia cells: trigger analyses. J Cell Sci. 1980 Dec;46:41–60. doi: 10.1242/jcs.46.1.41. [DOI] [PubMed] [Google Scholar]
  18. Matthews B. W. Determination of protein molecular weight, hydration, and packing from crystal density. Methods Enzymol. 1985;114:176–187. doi: 10.1016/0076-6879(85)14018-8. [DOI] [PubMed] [Google Scholar]
  19. Matthews B. W. Solvent content of protein crystals. J Mol Biol. 1968 Apr 28;33(2):491–497. doi: 10.1016/0022-2836(68)90205-2. [DOI] [PubMed] [Google Scholar]
  20. Peterson J. B., Heuser J. E., Nelson D. L. Dissociation and reassociation of trichocyst proteins: biochemical and ultrastructural studies. J Cell Sci. 1987 Feb;87(Pt 1):3–25. doi: 10.1242/jcs.87.1.3. [DOI] [PubMed] [Google Scholar]
  21. Pollack S., Steers E., Jr Thermal solubilization of the trichocysts from Paramecium aurelia. Exp Cell Res. 1973 Mar 30;78(1):186–190. doi: 10.1016/0014-4827(73)90053-0. [DOI] [PubMed] [Google Scholar]
  22. Rauh J. J., Nelson D. L. Calmodulin is a major component of extruded trichocysts from Paramecium tetraurelia. J Cell Biol. 1981 Dec;91(3 Pt 1):860–865. doi: 10.1083/jcb.91.3.860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ruiz F., Adoutte A., Rossignol M., Beisson J. Genetic analysis of morphogenetic processes in Paramecium. I. A mutation affecting trichocyst formation and nuclear division. Genet Res. 1976 Apr;27(2):109–122. doi: 10.1017/s0016672300016323. [DOI] [PubMed] [Google Scholar]
  24. Sardet C., Tardieu A., Luzzati V. Shape and size of bovine rhodopsin: a small-angle x-ray scattering study of a rhodopsin-detergent complex. J Mol Biol. 1976 Aug 15;105(3):383–407. doi: 10.1016/0022-2836(76)90100-5. [DOI] [PubMed] [Google Scholar]
  25. Schliwa M., van Blerkom J. Structural interaction of cytoskeletal components. J Cell Biol. 1981 Jul;90(1):222–235. doi: 10.1083/jcb.90.1.222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Steers E., Jr, Beisson J., Marchesi V. T. A structural protein extracted from the trichocyst of Paramecium aurelia. Exp Cell Res. 1969 Oct;57(2):392–396. doi: 10.1016/0014-4827(69)90165-7. [DOI] [PubMed] [Google Scholar]
  27. Tartakoff A. M. Perturbation of vesicular traffic with the carboxylic ionophore monensin. Cell. 1983 Apr;32(4):1026–1028. doi: 10.1016/0092-8674(83)90286-6. [DOI] [PubMed] [Google Scholar]
  28. Thomas J. O., Kornberg R. D. An octamer of histones in chromatin and free in solution. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2626–2630. doi: 10.1073/pnas.72.7.2626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tindall S. H. Selection of chemical spacers to improve isoelectric focusing resolving power: implications for use in two-dimensional electrophoresis. Anal Biochem. 1986 Dec;159(2):287–294. doi: 10.1016/0003-2697(86)90345-3. [DOI] [PubMed] [Google Scholar]
  30. Tooney N. M., Cohen C. Crystalline states of a modified fibrinogen. J Mol Biol. 1977 Feb 25;110(2):363–385. doi: 10.1016/s0022-2836(77)80077-6. [DOI] [PubMed] [Google Scholar]
  31. White S. P., Cohen C., Phillips G. N., Jr Structure of co-crystals of tropomyosin and troponin. 1987 Feb 26-Mar 4Nature. 325(6107):826–828. doi: 10.1038/325826a0. [DOI] [PubMed] [Google Scholar]
  32. Xia Z. X., Shamala N., Bethge P. H., Lim L. W., Bellamy H. D., Xuong N. H., Lederer F., Mathews F. S. Three-dimensional structure of flavocytochrome b2 from baker's yeast at 3.0-A resolution. Proc Natl Acad Sci U S A. 1987 May;84(9):2629–2633. doi: 10.1073/pnas.84.9.2629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. YUSA A. AN ELECTRON MICROSCOPE STUDY ON REGENERATION OF TRICHOCYSTS IN PARAMECIUM CAUDATUM. J Protozool. 1963 Aug;10:253–262. doi: 10.1111/j.1550-7408.1963.tb01673.x. [DOI] [PubMed] [Google Scholar]

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