Organic solvent extraction as a versatile procedure to identify hydrophobic chloroplast membrane proteins
| Autor: | Daphné Seigneurin-Berny, Myriam Ferro, Daniel Salvi, Jacques Joyard, Agnès Chapel, Norbert Rolland, Jérôme Garin |
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| Rok vydání: | 2000 |
| Předmět: |
Gel electrophoresis
Chloroplasts Chromatography Chemistry Detergents Clinical Biochemistry Peripheral membrane protein Membrane Proteins Biochemistry Mass Spectrometry Transmembrane protein Analytical Chemistry Membrane Solubility Membrane protein Thylakoid Proteome Solvents Electrophoresis Polyacrylamide Gel Salts Organic Chemicals Integral membrane protein |
| Zdroj: | Electrophoresis. 21:3517-3526 |
| ISSN: | 1522-2683 0173-0835 |
| DOI: | 10.1002/1522-2683(20001001)21:16<3517::aid-elps3517>3.0.co;2-h |
| Popis: | As a complementary approach to genome projects, proteomic analyses have been set up to identify new gene products. One of the major challenges in proteomics concerns membrane proteins, especially the minor ones. A procedure based on the differential extraction of membrane proteins in chloroform/methanol mixtures, was tested on the two different chloroplast membrane systems: envolope and thylakoid membranes. Combining the use of classical sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometry analyses, this procedure enabled identification of hydrophobic proteins. The propensity of hydrophobic proteins to partition in chloroform/methanol mixtures was directly correlated with the number of amino acid residues/number of putative transmembrane regions (Res/TM ratio). Regardless of the particular case of some lipid-interacting proteins, chloroform/methanol extractions allowed enrichment of hydrophobic proteins and exclusion of hydrophilic proteins from both membrane systems, thus demonstrating the versatility of the procedure. |
| Databáze: | OpenAIRE |
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