Identification and Origin of N-Linked β-D-N-Acetylglucosamine Monosaccharide Modifications on Arabidopsis Proteins
2012-11-08
Loading...
View/Download File
Persistent link to this item
Statistics
View StatisticsJournal Title
Journal ISSN
Volume Title
Title
Identification and Origin of N-Linked β-D-N-Acetylglucosamine Monosaccharide Modifications on Arabidopsis Proteins
Published Date
2012-11-08
Publisher
American Society of Plant Biologists
Type
Article
Abstract
Many plant proteins are modified with N-linked oligosaccharides at asparagine-X-serine/threonine sites during transit through the endoplasmic reticulum and the Golgi. We have identified a number of Arabidopsis (Arabidopsis thaliana) proteins with modifications consisting of an N-linked N-acetyl-D-glucosamine monosaccharide (N-GlcNAc). Electron transfer dissociation mass spectrometry analysis of peptides bearing this modification mapped the modification to asparagine-X-serine/threonine sites on proteins that are predicted to transit through the endoplasmic reticulum and Golgi. A mass labeling method was developed and used to study N-GlcNAc modification of two thioglucoside glucohydrolases (myrosinases), TGG1 and TGG2 (for thioglucoside glucohydrolase). These myrosinases are also modified with high-mannose (Man)-type glycans. We found that N-GlcNAc and high-Man-type glycans can occur at the same site. It has been hypothesized that N-GlcNAc modifications are generated when endo-β-N-acetylglucosaminidase (ENGase) cleaves N-linked glycans. We examined the effects of mutations affecting the two known Arabidopsis ENGases on N-GlcNAc modification of myrosinase and found that modification of TGG2 was greatly reduced in one of the single mutants and absent in the double mutant. Surprisingly, N-GlcNAc modification of TGG1 was not affected in any of the mutants. These data support the hypothesis that ENGases hydrolyze high-Man glycans to produce some of the N-GlcNAc modifications but also suggest that some N-GlcNAc modifications are generated by another mechanism. Since N-GlcNAc modification was detected at only one site on each myrosinase, the production of the N-GlcNAc modification may be regulated.
Keywords
Description
Related to
Replaces
License
Collections
Series/Report Number
Funding information
Isbn identifier
Doi identifier
Previously Published Citation
Young-Cheon Kim, Neal Jahren, Matthew D. Stone, Namrata D. Udeshi, Todd W. Markowski, Bruce A. Witthuhn, Jeffrey Shabanowitz, Donald F. Hunt, and Neil E. Olszewski. Identification and Origin of N-Linked β-D-N-Acetylglucosamine Monosaccharide Modifications on Arabidopsis Proteins. Plant Physiology. 2013 161: 455-464. First Published on November 8, 2012; doi:10.1104/pp.112.208900
Suggested citation
Olszewski, Neil E.; Kim, Young-Cheon; Jahren, Neal; Stone, Mathew D.; Udeshi, Namrata D.; Markowski, Todd W.; Witthuhn, Bruce A.; Shabanowitz, Jeffrey; Huntand, Donald F.. (2012). Identification and Origin of N-Linked β-D-N-Acetylglucosamine Monosaccharide Modifications on Arabidopsis Proteins. Retrieved from the University Digital Conservancy, https://hdl.handle.net/11299/143832.
Content distributed via the University Digital Conservancy may be subject to additional license and use restrictions applied by the depositor. By using these files, users agree to the Terms of Use. Materials in the UDC may contain content that is disturbing and/or harmful. For more information, please see our statement on harmful content in digital repositories.