Effectiveness and limitation of two-dimensional gel electrophoresis in bacterial membrane protein proteomics and perspectives
| Autor: | Keigo Bunai, Kunio Yamane |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
Proteomics
Two-dimensional gel electrophoresis Chromatography Proteome Isoelectric focusing Chemistry Clinical Biochemistry Membrane Proteins Cell Biology General Medicine Biochemistry Transmembrane protein Analytical Chemistry Electrophoresis Membrane protein Isotope Labeling Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Electrophoresis Gel Two-Dimensional Isoelectric Focusing Polyacrylamide gel electrophoresis Genome Bacterial Bacillus subtilis |
| Zdroj: | Journal of Chromatography B. 815:227-236 |
| ISSN: | 1570-0232 |
| Popis: | Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) using isoelectric focusing and SDS-PAGE in the first and second dimensions, respectively, is an established means of simultaneously separating over 1000 proteins and two new types have recently been developed. These procedures have significant shortcomings such as low load ability and poor separation of hydrophobic, acidic and alkaline proteins. We therefore modified the protocols to analyze the Bacillus subtilis membrane proteome. The 2D-PAGE techniques effectively separated membrane proteins having one and two transmembrane segments but not those with more than four. Compared with new LC/MS/MS procedures that are independent of electrophoretic separation, 2D-PAGE can globally analyze and quantify proteins at various stages of the cell cycle when labeled with isotopes such as 35S-methionine or the stable isotope, 15N. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect