Resolving and identifying protein components of plant mitochondrial respiratory complexes using three dimensions of gel electrophoresis.
Analyzing highly hydrophobic proteins is a challenge for identification protocols based on gel separation and mass spectrometry. We combined Blue Native and 2D tricine gel electrophoresis to allow separation and identification of respiratory complex subunits from Arabidopsis mitochondria. We identified many of the highly hydrophobic mitochondrion-encoded subunits (GRAVY scores between +0.6 to +1.4) and also found a number of nucleus-encoded proteins associated with complex I for the first time in plants.
Meyer
Etienne H
EH
ARC Center of Excellence in Plant Energy Biology, The University of Western Australia, 35 Stirling Highway, Crawley WA 6009, Australia.
Taylor
Nicolas L
NL
Millar
A Harvey
AH
D000595:Amino Acid Sequence; D000818:Animals; D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D002417:Cattle; D019069:Cell Respiration / Q000502:physiology; D042967:Electron Transport Complex I / Q000737:chemistry / Q000302:isolation & purification; D014450:Electron Transport Complex III / Q000737:chemistry / Q000302:isolation & purification; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D008928:Mitochondria / Q000737:chemistry / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D021241:Spectrometry, Mass, Electrospray Ionization
D029681:Arabidopsis Proteins; D024101:Mitochondrial Proteins; D042967:Electron Transport Complex I; D014450:Electron Transport Complex III