Mapping the Arabidopsis organelle proteome.
A challenging task in the study of the secretory pathway is the identification and localization of new proteins to increase our understanding of the functions of different organelles. Previous proteomic studies of the endomembrane system have been hindered by contaminating proteins, making it impossible to assign proteins to organelles. Here we have used the localization of organelle proteins by the isotope tagging technique in conjunction with isotope tags for relative and absolute quantitation and 2D liquid chromatography for the simultaneous assignment of proteins to multiple subcellular compartments. With this approach, the density gradient distributions of 689 proteins from Arabidopsis thaliana were determined, enabling confident and simultaneous localization of 527 proteins to the endoplasmic reticulum, Golgi apparatus, vacuolar membrane, plasma membrane, or mitochondria and plastids. This parallel analysis of endomembrane components has enabled protein steady-state distributions to be determined. Consequently, genuine organelle residents have been distinguished from contaminating proteins and proteins in transit through the secretory pathway.
Dunkley
Tom P J
TP
Department of Biochemistry, University of Cambridge, Downing Site, Cambridge CB2 1QW, United Kingdom.
Hester
Svenja
S
Shadforth
Ian P
IP
Runions
John
J
Weimar
Thilo
T
Hanton
Sally L
SL
Griffin
Julian L
JL
Bessant
Conrad
C
Brandizzi
Federica
F
Hawes
Chris
C
Watson
Rod B
RB
Dupree
Paul
P
Lilley
Kathryn S
KS
D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D015388:Organelles / Q000235:genetics / Q000378:metabolism; D010449:Peptide Mapping; D020543:Proteome / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism
D029681:Arabidopsis Proteins; D020543:Proteome; D011993:Recombinant Fusion Proteins