Isolation and Solubilization of Cellular Membrane Proteins from Bacteria

作者: Kheir Zuobi-Hasona , L. Jeannine Brady

DOI: 10.1007/978-1-60327-210-0_23

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摘要: Membrane proteins are rarely identified in two-dimensional electrophoretic (2-DE) proteomics maps. This is because of low abundance, poor solubility, and inherent hydrophobicity. In this study, membrane preparations from the Gram-positive bacterium Streptococcus mutans were isolated protoplasts by mechanical grinding. extracted using a mixture trifluroethanol chloroform, solubilized highly chaotropic buffer containing ASB-14 Triton X-100 subjected to gel electrophoresis.

参考文章(18)
Valerie C. Wasinger, J. Dennis Pollack, Ian Humphery-Smith, The proteome of Mycoplasma genitalium European Journal of Biochemistry. ,vol. 267, pp. 1571- 1582 ,(2000) , 10.1046/J.1432-1327.2000.01183.X
Mark P. Molloy, Ben R. Herbert, Martin B. Slade, Thierry Rabilloud, Amanda S. Nouwens, Keith L. Williams, Andrew A. Gooley, Proteomic analysis of the Escherichia coli outer membrane European Journal of Biochemistry. ,vol. 267, pp. 2871- 2881 ,(2000) , 10.1046/J.1432-1327.2000.01296.X
C. Bordier, Phase separation of integral membrane proteins in Triton X-114 solution. Journal of Biological Chemistry. ,vol. 256, pp. 1604- 1607 ,(1981) , 10.1016/S0021-9258(19)69848-0
Thierry Rabilloud, Thierry Blisnick, Manfred Heller, Sylvie Luche, Ruedi Aebersold, Joël Lunardi, Catherine Braun-Breton, Analysis of membrane proteins by two-dimensional electrophoresis: comparison of the proteins extracted from normal or Plasmodium falciparum-infected erythrocyte ghosts. Electrophoresis. ,vol. 20, pp. 3603- 3610 ,(1999) , 10.1002/(SICI)1522-2683(19991201)20:18<3603::AID-ELPS3603>3.0.CO;2-V
Véronique Santoni, Mark Molloy, Thierry Rabilloud, Membrane proteins and proteomics: un amour impossible? Electrophoresis. ,vol. 21, pp. 1054- 1070 ,(2000) , 10.1002/(SICI)1522-2683(20000401)21:6<1054::AID-ELPS1054>3.0.CO;2-8
Jacques M. P. Deshusses, Jennifer A. Burgess, Alexander Scherl, Yvan Wenger, Nadia Walter, Véronique Converset, Salvo Paesano, Garry L. Corthals, Denis F. Hochstrasser, Jean-Charles Sanchez, Exploitation of specific properties of trifluoroethanol for extraction and separation of membrane proteins. Proteomics. ,vol. 3, pp. 1418- 1424 ,(2003) , 10.1002/PMIC.200300492
Mark P. Molloy, Ben R. Herbert, Bradley J. Walsh, Margaret I. Tyler, Mathew Traini, Jean-Charles Sanchez, Denis F. Hochstrasser, Keith L. Williams, Andrew A. Gooley, Extraction of membrane proteins by differential solubilization for separation using two‐dimensional gel electrophoresis Electrophoresis. ,vol. 19, pp. 837- 844 ,(1998) , 10.1002/ELPS.1150190539
Mark P. Molloy, Ben R. Herbert, Keith L. Williams, Andrew A. Gooley, Extraction of Escherichia coli proteins with organic solvents prior to two‐dimensional electrophoresis Electrophoresis. ,vol. 20, pp. 701- 704 ,(1999) , 10.1002/(SICI)1522-2683(19990101)20:4/5<701::AID-ELPS701>3.0.CO;2-5