The hydrophobic proteome of mitochondrial membranes from Arabidopsis cell suspensions.
The development of mitochondria and the integration of their function within a plant cell rely on the presence of a complex biochemical machinery located within their limiting membranes. The aim of the present work was: (1) to enhance our understanding of the biochemical machinery of mitochondrial membranes and (2) to test the versatility of the procedure developed for the identification of the hydrophobic proteome of the chloroplast envelope [Molecular and Cellular Proteomics 2 (2003) 325-345]. A proteomic analysis was performed, to provide the most exhaustive view of the protein repertoire of these membranes. For this purpose, highly purified mitochondria were prepared from Arabidopsis cultured cells and membrane proteins were extracted. To get a more exhaustive array of membrane proteins from Arabidopsis mitochondria, from the most to the less hydrophobic ones, various extraction procedures (chloroform/methanol extraction, alkaline or saline treatments) were applied. LC-MS/MS analyses were then performed on each membrane subfraction, leading to the identification of more than 110 proteins. The identification of these proteins is discussed with respect to their mitochondrial localization, their physicochemical properties and their implications in the metabolism of mitochondria. In order to provide a new overview of the biochemical machinery of the plant mitochondria, proteins identified during this work were compared to the lists of proteins identified during previous proteomic analyses performed on plant and algae mitochondria (Arabidopsis, pea, Chlamydomonas, rice, etc.). A total of 502 proteins are listed. About 40% of the 114 proteins identified during this work were not identified during previous proteomic studies performed on mitochondria.
Laboratoire de Chimie des Protéines, ERM-0201 INSERM/CEA, France.
Sabine
S
Brugière
Solène
S
Kowalski
Myriam
M
Ferro
Daphné
D
Seigneurin-Berny
Stéphane
S
Miras
Daniel
D
Salvi
Stéphane
S
Ravanel
Pierre
P
d'Hérin
Jérôme
J
Garin
Jacques
J
Bourguignon
Jacques
J
Joyard
Norbert
N
Rolland
D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000032:analysis / Q000302:isolation & purification; D002478:Cells, Cultured; D002736:Chloroplasts / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism*; D008565:Membrane Proteins / Q000032:analysis* / Q000302:isolation & purification; D008928:Mitochondria / Q000378:metabolism*; D020543:Proteome / Q000032:analysis* / Q000302:isolation & purification
D029681:Arabidopsis Proteins; D008565:Membrane Proteins; D020543:Proteome