Spheroplast

Spheroplast

A spheroplast is a cell from which the cell wall has been almost completely removed, as by the action of penicillin. The name stems from the fact that after a microbe's cell wall is digested, membrane tension causes the cell to acquire a characteristic spherical shape. Spheroplasts are osmotically fragile, and will lyse if transferred to a hypotonic solution.

Uses and applications

Patch clamping

Specially prepared giant spheroplasts of Gram-negative bacteria can be used to study the function of bacterial ion channels through a technique called patch clamp, which was originally designed for characterizing the behavior of neurons and other excitable cells. To prepare giant spheroplasts, bacteria are grown in a medium containing chemicals that prevent the cells from dividing completely. This causes bacteria to form long "snakes" that share a single membrane and cytoplasm. After a period of time, the cell walls of the "snakes" are digested, and the bacteria collapse into very large spheres surrounded by a single lipid bilayer. The membrane can then be analyzed on a patch clamp apparatus to determine the phenotype of the ion channels embedded in it. It is also common to overexpress a particular channel to amplify its effect and make it easier to characterize.

The technique of patch clamping giant "E. coli" spheroplast have been used extensively for studying the native mechanosensitive channels (MscL, MscS, and MscM) of "E. coli" since 1987. [Martinac, B., Buechner, M., Delcour, A. H., Adler, J., and Kung, C. (1987) Pressure-sensitive ion channel in Escherichia coli. Proc. Natl. Acad. Sci. USA 84, 2297-2301.] [Blount, P., Sukharev, S. I., Moe, P. C., Martinac, B., and Kung, C. (1999) Mechanosensitive channels of bacteria. Methods in Enzymology 294, 458-482.] Recently, it has been extended to study other heterologously expressed ion channels and has been shown that the giant "E. coli" shperoplast can be used as an ion-channel expression system comparable to Xenopus oocyte. [ Santos, J. S., Lundby, A., Zazueta, C., and Montal, M. (2006) Molecular template for a voltage sensor in a novel K+ channel. I. Identification and functional characterization of KvLm, a voltage-gated K+ channel from Listeria monocytogenes. Journal of General Physiology 128(3), 283-292.] [Nakayama, Y., Fujiu, K., Sokabe, M., and Yoshimura, K. (2007) Molecular and electrophysiological characterization of a mechanosensitive channel expressed in the chloroplasts of Chlamydomonas. Proc. Natl. Acad. Sci. USA 104, 5883-5888.] [Kuo, M. M.-C., Baker, K. A., Wong, L., and Choe, S. (2007) Dynamic oligomeric conversions of the cytoplasmic RCK domains mediate MthK potassium channel activity. Proc. Natl. Acad. Sci. USA 104, 2151-2156.] [Kuo, M. M.-C., Saimi, Y., Kung, C., and Choe, S. (2007). Patch-clamp and phenotypic analyses of a prokaryotic cyclic nucleotide-gated K+ channel using Escherichia Coli as a host. J. Biol. Chem. 282, 24294-24301.]

Cell lysis

Yeast cells are normally protected by a thick cell wall which makes extraction of cellular proteins difficult. Enzymatic digestion of the cell wall with zymolyase, creating spheroplasts, renders the cells vulnerable to easy lysis with detergents or rapid osmolar pressure changes.

References

External links

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Look at other dictionaries:

  • spheroplast — spheroplast. См. сферопласт. (Источник: «Англо русский толковый словарь генетических терминов». Арефьев В.А., Лисовенко Л.А., Москва: Изд во ВНИРО, 1995 г.) …   Молекулярная биология и генетика. Толковый словарь.

  • spheroplast — (AlteRNAtive spelling: sphaeroplast). A microbial or plant cell from which most of the cell wall has been removed, usually by enzymatic treatment. Strictly, in a spheroplast, some of the cell wall remains, while in a protoplast the cell wall has… …   Glossary of Biotechnology

  • spheroplast — noun Date: circa 1920 a bacterium or yeast cell that is modified (as by enzymatic action) so that there is partial loss of the cell wall and increased osmotic sensitivity …   New Collegiate Dictionary

  • spheroplast — (sfĭr o plast) A relatively spherical cell formed by the weakening or partial removal of the rigid cell wall component (e.g., by penicillin treatment of gram negative bacteria). Spheroplasts are usually osmotically sensitive …   Dictionary of microbiology

  • spheroplast — Bacterium from which the cell wall has been removed but that has not lysed …   Dictionary of molecular biology

  • spheroplast — /sfear euh plast , sfer /, n. Bacteriol. a Gram negative bacterial cell with a cell wall that has been altered or is partly missing, resulting in a spherical shape. [1915 20; SPHERO + PLAST] * * * …   Universalium

  • spheroplast — noun A cell from which the cell wall has been removed …   Wiktionary

  • spheroplast — A bacterial cell from which the rigid cell wall has been incompletely removed. The bacterium loses its characteristic shape and becomes round. [sphero + G. plastos, formed] * * * sphe·ro·plast sfir ə .plast, sfer n a bacterium or yeast cell that… …   Medical dictionary

  • spheroplast — sphe·ro·plast …   English syllables

  • spheroplast — sphe•ro•plast [[t]ˈsfɪər əˌplæst, ˈsfɛr [/t]] n. mcr a Gram negative bacterial cell with a cell wall that has been altered or is partly missing, resulting in a spherical shape • Etymology: 1958 …   From formal English to slang

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