Proteomic analysis reveals O-GlcNAc modification on proteins with key regulatory functions inArabidopsis.
Genetic studies have shown essential functions of O-linkedN-acetylglucosamine (O-GlcNAc) modification in plants. However, the proteins and sites subject to this posttranslational modification are largely unknown. Here, we report a large-scale proteomic identification of O-GlcNAc-modified proteins and sites in the model plantArabidopsis thalianaUsing lectin weak affinity chromatography to enrich modified peptides, followed by mass spectrometry, we identified 971 O-GlcNAc-modified peptides belonging to 262 proteins. The modified proteins are involved in cellular regulatory processes, including transcription, translation, epigenetic gene regulation, and signal transduction. Many proteins have functions in developmental and physiological processes specific to plants, such as hormone responses and flower development. Mass spectrometric analysis of phosphopeptides from the same samples showed that a large number of peptides could be modified by either O-GlcNAcylation or phosphorylation, but cooccurrence of the two modifications in the same peptide molecule was rare. Our study generates a snapshot of the O-GlcNAc modification landscape in plants, indicating functions in many cellular regulation pathways and providing a powerful resource for further dissecting these functions at the molecular level.
Department of Plant Biology, Carnegie Institution for Science, Stanford, CA 94305.|Department of Pharmaceutical Chemistry, University of California, San Francisco, CA 94158.
Shou-Ling
SL
Xu
Department of Pharmaceutical Chemistry, University of California, San Francisco, CA 94158.
Robert J
RJ
Chalkley
Department of Pharmaceutical Chemistry, University of California, San Francisco, CA 94158.
Jason C
JC
Maynard
Basic Forestry and Proteomics Research Center, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
Wenfei
W
Wang
Department of Plant and Microbial Biology, University of California, Berkeley, CA, 94720.|Plant Gene Expression Center, United States Department of Agriculture/Agriculture Research Service, Albany, CA94710.
Weimin
W
Ni
Thermo Fisher Scientific, San Jose, CA 95134.
Xiaoyue
X
Jiang
Basic Forestry and Proteomics Research Center, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
Kihye
K
Shin
Basic Forestry and Proteomics Research Center, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
Ling
L
Cheng
Department of Plant Biology, Carnegie Institution for Science, Stanford, CA 94305.
Dasha
D
Savage
Thermo Fisher Scientific, San Jose, CA 95134.
Andreas F R
AF
Hühmer
Department of Pharmaceutical Chemistry, University of California, San Francisco, CA 94158; zywang24@stanford.edu alb@cgl.ucsf.edu.
Alma L
AL
Burlingame
Department of Plant Biology, Carnegie Institution for Science, Stanford, CA 94305; zywang24@stanford.edu alb@cgl.ucsf.edu.
Zhi-Yong
ZY
Wang
D000117:Acetylglucosamine / Q000378:metabolism*; D000215:Acylation; D017360:Arabidopsis / Q000502:physiology*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D042002:Chromatin Assembly and Disassembly / Q000502:physiology; D002846:Chromatography, Affinity; D035264:Flowers / Q000254:growth & development; D006031:Glycosylation; D037102:Lectins / Q000737:chemistry; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D040901:Proteomics / Q000379:methods; D015398:Signal Transduction / Q000502:physiology*; D053719:Tandem Mass Spectrometry
D029681:Arabidopsis Proteins; D037102:Lectins; D000117:Acetylglucosamine
Arabidopsis; O-GlcNAcylation; phosphorylation; plant; proteomics