Spheroplast 496297 202208006 2008-03-31T02:34:13Z Sabedon 2083035 /* External links */ added [[Category:Microbiology]] [[Image:Patchclamp Spheroplast1.jpg|thumb|right|An ''E.coli'' [[spheroplast]] patched with a glass pipette.]]<!-- Image with unknown copyright status removed: [[Image:Spheroplast.JPG|thumb|right|Top left, growing E. coli cells; top right, filamentous E coli cells; bottom left, a glass pipette attached to a giant E coli spheroplast; bottom right, a small membrane patch is drawn into the tip of the glass pipette to form a seal, allowing patch clamp analysis of channels in the membrane patch. Scale bars are 10 micrometers long.]] --> 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.<ref>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.</ref><ref>Blount, P., Sukharev, S. I., Moe, P. C., Martinac, B., and Kung, C. (1999) Mechanosensitive channels of bacteria. Methods in Enzymology 294, 458-482.</ref> 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.<ref> 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.</ref><ref>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.</ref><ref>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.</ref><ref>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.</ref> === 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== <references/> ==External links== * {{MeshName|Spheroplasts}} [[Category:Bacteria]] [[Category:Microbiology]] [[ja:スフェロプラスト]]