| O-GlcNAcylation of the Plum pox virus capsid protein catalyzed by SECRET AGENT: characterization of O-GlcNAc sites by electron transfer dissociation mass spectrometry. | |
| | |
MedLine Citation:
|
PMID: 20676902 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
The capsid protein of Plum pox virus (PPV-CP) is modified with O-linked β-N-acetylglucosamine (O-GlcNAc). In Arabidopsis thaliana this modification is made by an O-GlcNAc transferase named SECRET AGENT (SEC). Modification of PPV-CP by SEC is hypothesized to have a direct role in the infection process, because virus titer and rate of spread are reduced in SEC mutants. Previous studies used deletion mapping and site-directed mutagenesis to identify four O-GlcNAc sites on the capsid protein that are modified by Escherichia coli-expressed SEC. The infection process was not affected when two of these sites were mutated suggesting that O-GlcNAcylation of these sites does not have a significant role in the infection process or that a subset of the modifications is sufficient. Since it is possible that the mutational mapping approach missed or incorrectly identified O-GlcNAc sites, the modifications produced by E. coli-expressed SEC were characterized using mass spectrometry. O-GlcNAcylated peptides were enzymatically tagged with galactose, the products were enriched on immobilized Ricinus communis agglutinin I and sequenced by electron transfer dissociation (ETD) mass spectrometry. Five O-GlcNAc sites on PPV-CP were identified. Two of these sites were not identified in by the previous mutational mapping. In addition, one site previously predicted by mutation mapping was not detected, but modification of this site was not supported when the mutation mapping was repeated. This study suggests that mapping modification sites by ETD mass spectrometry is more comprehensive and accurate than mutational mapping. |
| | |
Authors:
|
Young-Cheon Kim; Namrata D Udeshi; Jeremy L Balsbaugh; Jeffrey Shabanowitz; Donald F Hunt; Neil E Olszewski |
Publication Detail:
|
Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S. Date: 2010-07-31 |
Journal Detail:
|
Title: Amino acids Volume: 40 ISSN: 1438-2199 ISO Abbreviation: Amino Acids Publication Date: 2011 Mar |
Date Detail:
|
Created Date: 2011-02-18 Completed Date: 2011-05-24 Revised Date: 2011-09-26 |
Medline Journal Info:
|
Nlm Unique ID: 9200312 Medline TA: Amino Acids Country: Austria |
Other Details:
|
Languages: eng Pagination: 869-76 Citation Subset: IM |
Affiliation:
|
Department of Plant Biology, Microbial and Plant Genomics Institute, 1445 Gortner Ave., St. Paul, MN 55108, USA. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Acetylglucosamine
/
metabolism* Amino Acid Sequence Arabidopsis Proteins / genetics, metabolism* Biocatalysis Capsid Proteins / chemistry*, genetics, metabolism* Glycosylation Mass Spectrometry Molecular Sequence Data N-Acetylglucosaminyltransferases / genetics, metabolism* Peptide Mapping Plum Pox Virus / chemistry, genetics, metabolism* |
| Grant Support | |
ID/Acronym/Agency:
|
GM37537/GM/NIGMS NIH HHS; R01 GM037537-25/GM/NIGMS NIH HHS |
| Chemical | |
Reg. No./Substance:
|
0/Arabidopsis Proteins; 0/Capsid Proteins; 7512-17-6/Acetylglucosamine; EC 2.4.1.-/N-Acetylglucosaminyltransferases; EC 2.4.1.-/SECRET AGENT protein, Arabidopsis |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Diacyl glycerol arginine-based surfactants: biological and physicochemical properties of catanionic ...
Next Document: Plasma free amino acid kinetics in rainbow trout (Oncorhynchus mykiss) using a bolus injection of 15...