A comprehensive two-dimensional map of cytosolic proteins ofBacillus subtilis
2001; Wiley; Volume: 22; Issue: 14 Linguagem: Inglês
10.1002/1522-2683(200108)22
ISSN1522-2683
AutoresKnut Büttner, Jörg Bernhardt, Christian Scharf, Roland Schmid, Ulrike Mäder, Christine Eymann, Haike Antelmann, Andrea Völker, Uwe Völker, Michael Hecker,
Tópico(s)Bacterial Genetics and Biotechnology
ResumoELECTROPHORESISVolume 22, Issue 14 p. 2908-2935 Research Article A comprehensive two-dimensional map of cytosolic proteins of Bacillus subtilis Knut Büttner, Knut Büttner Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität Greifswald, GermanySearch for more papers by this authorJörg Bernhardt, Jörg Bernhardt Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität Greifswald, GermanySearch for more papers by this authorChristian Scharf, Christian Scharf Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität Greifswald, GermanySearch for more papers by this authorRoland Schmid, Roland Schmid Abteitung für Mikrobiologie, Universität Osnabrück GermanySearch for more papers by this authorUlrike Mäder, Ulrike Mäder Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität Greifswald, GermanySearch for more papers by this authorChristine Eymann, Christine Eymann Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität Greifswald, GermanySearch for more papers by this authorHeike Antelmann, Heike Antelmann Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität Greifswald, GermanySearch for more papers by this authorAndrea Völker, Andrea Völker Laboratorium für Mikrobiologie, Philipps-Universität und Max-Planck-Institut für terrestrische Mikrobiologie, Marburg, GermanySearch for more papers by this authorUwe Völker, Uwe Völker Laboratorium für Mikrobiologie, Philipps-Universität und Max-Planck-Institut für terrestrische Mikrobiologie, Marburg, GermanySearch for more papers by this authorMichael Hecker, Corresponding Author Michael Hecker [email protected] Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität Greifswald, GermanyErnst-Moritz-Arndt-Universität Greifswald, Institut für Mikrobiologie, Friedrich-Ludwig-Jahn-Straße 15, D-17487 Greifswald, Germany Fax: +49-(0)-3834-864202===Search for more papers by this author Knut Büttner, Knut Büttner Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität Greifswald, GermanySearch for more papers by this authorJörg Bernhardt, Jörg Bernhardt Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität Greifswald, GermanySearch for more papers by this authorChristian Scharf, Christian Scharf Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität Greifswald, GermanySearch for more papers by this authorRoland Schmid, Roland Schmid Abteitung für Mikrobiologie, Universität Osnabrück GermanySearch for more papers by this authorUlrike Mäder, Ulrike Mäder Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität Greifswald, GermanySearch for more papers by this authorChristine Eymann, Christine Eymann Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität Greifswald, GermanySearch for more papers by this authorHeike Antelmann, Heike Antelmann Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität Greifswald, GermanySearch for more papers by this authorAndrea Völker, Andrea Völker Laboratorium für Mikrobiologie, Philipps-Universität und Max-Planck-Institut für terrestrische Mikrobiologie, Marburg, GermanySearch for more papers by this authorUwe Völker, Uwe Völker Laboratorium für Mikrobiologie, Philipps-Universität und Max-Planck-Institut für terrestrische Mikrobiologie, Marburg, GermanySearch for more papers by this authorMichael Hecker, Corresponding Author Michael Hecker [email protected] Institut für Mikrobiologie, Ernst-Moritz-Arndt-Universität Greifswald, GermanyErnst-Moritz-Arndt-Universität Greifswald, Institut für Mikrobiologie, Friedrich-Ludwig-Jahn-Straße 15, D-17487 Greifswald, Germany Fax: +49-(0)-3834-864202===Search for more papers by this author First published: 23 August 2001 https://doi.org/10.1002/1522-2683(200108)22:14 3.0.CO;2-MCitations: 196AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat Abstract Proteomics relying on two-dimensional (2-D) gel electrophoresis of proteins followed by spot identification with mass spectrometry is an excellent experimental tool for physiological studies opening a new perspective for understanding overall cell physiology. This is the intriguing outcome of a method introduced by Klose and O'Farrell independently 25 years ago. Physiological proteomics requires a 2-D reference map on which most of the main proteins were identified. In this paper, we present such a reference map with more than 300 entries for Bacillus subtilis proteins with an isoelectric point (pI) between 4 and 7. The most abundant proteins of exponentially growing cells were compiled and shown to perform mainly housekeeping functions in glycolysis, tricarboxylic acid cycle (TCC), amino acid biosynthesis and translation as well as protein quality control. Furthermore, putative post-translational modifications were shown at a large scale, with 47 proteins in total forming more than one spot. In a few selected cases evidence for phosphorylation of these proteins is presented. The proteome analysis in the standard pI range was complemented by either stretching the most crowded regions in a narrow pH gradient 4.5–5.5, or by adding other fractions of the total B. subtilis proteome such as alkaline proteins as well as extracellular proteins. A big challenge for future studies is to provide an experimental protocol covering the fraction of intrinsic membrane proteins that almost totally escaped detection by the experimental procedure used in this study.* References 1 Kunst, F., et al., Nature 1997, 390, 249–256. 2 Klose, J., Humangenetik 1975, 26, 231–243. 3 O'Farrell, P. H., J. Biol. Chem. 1975, 250, 4007–4021. 4 Richter, A., Hecker, M., FEMS Microbiol. Lett. 1986, 36, 69–71. 5 Hecker, M., Richter, A., Schroeter, A., Wölfel, L., Mach, F., Z. 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