Organellar proteomics reveals Golgi arginine dimethylation.
The Golgi complex functions to posttranslationally modify newly synthesized proteins and lipids and to sort them to their sites of function. In this study, a stacked Golgi fraction was isolated by classical cell fractionation, and the protein complement (the Golgi proteome) was characterized using multidimensional protein identification technology. Many of the proteins identified are known residents of the Golgi, and 64% of these are predicted transmembrane proteins. Proteins localized to other organelles also were identified, strengthening reports of functional interfacing between the Golgi and the endoplasmic reticulum and cytoskeleton. Importantly, 41 proteins of unknown function were identified. Two were selected for further analysis, and Golgi localization was confirmed. One of these, a putative methyltransferase, was shown to be arginine dimethylated, and upon further proteomic analysis, arginine dimethylation was identified on 18 total proteins in the Golgi proteome. This survey illustrates the utility of proteomics in the discovery of novel organellar functions and resulted in 1) a protein profile of an enriched Golgi fraction; 2) identification of 41 previously uncharacterized proteins, two with confirmed Golgi localization; 3) the identification of arginine dimethylated residues in Golgi proteins; and 4) a confirmation of methyltransferase activity within the Golgi fraction.
Wu
Christine C
CC
Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
MacCoss
Michael J
MJ
Mardones
Gonzalo
G
Finnigan
Claire
C
Mogelsvang
Soren
S
Yates
John R
JR
Howell
Kathryn E
KE
D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000378:metabolism*; D002458:Cell Fractionation; D004721:Endoplasmic Reticulum / Q000378:metabolism; D020869:Gene Expression Profiling; D006056:Golgi Apparatus / Q000378:metabolism*; D008745:Methylation; D008969:Molecular Sequence Data; D021381:Protein Transport; D011506:Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics; D051381:Rats
D011506:Proteins; D001120:Arginine