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Organic solvent extraction as a versatile procedure to identify hydrophobic chloroplast membrane proteins
Author(s) -
Ferro Myriam,
SeigneurinBerny Daphné,
Rolland Norbert,
Chapel Agnès,
Salvi Daniel,
Garin Jérome,
Joyard Jacques
Publication year - 2000
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(20001001)21:16<3517::aid-elps3517>3.0.co;2-h
Subject(s) - membrane protein , chemistry , chromatography , chloroform , membrane , transmembrane protein , gel electrophoresis , sodium dodecyl sulfate , biochemistry , thylakoid , proteomics , chloroplast , gene , receptor
Abstract 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.