Proteomic study of the Arabidopsis thaliana chloroplastic envelope membrane utilizing alternatives to traditional two-dimensional electrophoresis.
With the completion of the sequencing of the Arabidopsis genome and with the significant increase in the amount of other plant genome and expressed sequence tags (ESTs) data, plant proteomics is rapidly becoming a very active field. We have pursued a high-throughput mass spectrometry-based proteomics approach to identify and characterize membrane proteins localized to the Arabidopsis thaliana chloroplastic envelope membrane. In this study, chloroplasts were prepared from plate- or soil-grown Arabidopsis plants using a novel isolation procedure, and "mixed" envelopes were subsequently isolated using sucrose step gradients. We applied two alternative methodologies, off-line multidimensional protein identification technology (Off-line MUDPIT) and one-dimensional (1D) gel electrophoresis followed by proteolytic digestion and liquid chromatography coupled with tandem mass spectrometry (Gel-C-MS/MS), to identify envelope membrane proteins. This proteomic study enabled us to identify 392 nonredundant proteins.
Froehlich
John E
JE
Department of Energy Plant Research Laboratory, Michigan Proteome Consortium, Michigan State University, East Lansing, Michigan 48824, USA.
Wilkerson
Curtis G
CG
Ray
W Keith
WK
McAndrew
Rosemary S
RS
Osteryoung
Katherine W
KW
Gage
Douglas A
DA
Phinney
Brett S
BS
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D029681:Arabidopsis Proteins / Q000032:analysis* / Q000145:classification / Q000302:isolation & purification; D015153:Blotting, Western; D002736:Chloroplasts / Q000737:chemistry* / Q000502:physiology; D019295:Computational Biology; D030562:Databases, Protein; D001330:Electronic Data Processing; D004591:Electrophoresis, Polyacrylamide Gel; D008401:Gas Chromatography-Mass Spectrometry; D008565:Membrane Proteins / Q000032:analysis* / Q000145:classification / Q000302:isolation & purification; D008969:Molecular Sequence Data; D040901:Proteomics
D029681:Arabidopsis Proteins; D008565:Membrane Proteins