Two-dimensional electrophoretic analysis ofCorynebacterium glutamicum membrane fraction and surface proteins
| Autor: | Andreas Burkovski, Reinhard Krämer, Gregor Wersch, Melanie Finkemeier, Walter Pfefferle, Thomas Hermann |
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| Rok vydání: | 2000 |
| Předmět: |
Gel electrophoresis
Chromatography Isoelectric focusing Blotting Western Molecular Sequence Data Clinical Biochemistry Corynebacterium Membrane Proteins Biology biology.organism_classification Biochemistry Analytical Chemistry Corynebacterium glutamicum Electrophoresis Bacterial Proteins Membrane protein Cytoplasm Electrophoresis Gel Two-Dimensional Amino Acid Sequence Peptide sequence |
| Zdroj: | Electrophoresis. 21:654-659 |
| ISSN: | 1522-2683 0173-0835 |
| DOI: | 10.1002/(sici)1522-2683(20000201)21:3<654::aid-elps654>3.0.co;2-1 |
| Popis: | An improved protocol for the two-dimensional analysis of proteins of the Corynebacterium glutamicum cytoplasmic membrane fraction is described. By use of increased 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) concentrations (2-4%) and an optimized electrophoresis protocol, horizontal streaking of proteins of the cytoplasmic membrane fraction was almost completely avoided. More important, in contrast to a previously published method, both a sample tray and IPG-phor isoelectric focusing unit can be used for the in-gel application of proteins. The described protocol was also found to be suitable for hydrophilic cytoplasmic proteins. Additionally, the preparation and analysis of C. glutamicum cell surface proteins is described. Proteins were extracted with lauroyl sarcosinate and 100-120 spots were separated on two-dimensional (2-D) gels in comparison to 18-20 spots observed previously by standard sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). C. glutamicum proteins can now be separated into three distinct fractions resembling different functional units of the bacterial cell. |
| Databáze: | OpenAIRE |
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