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An Isotope-Coded Photocleavable Probe for Quantitative Profiling of Protein O-GlcNAcylation

Li, Jingchao and Li, Zhonghua and Duan, Xiaotao and Qin, Ke and Dang, Liuyi and Sun, Shisheng and Cai, Li and Hsieh-Wilson, Linda C. and Wu, Liming and Yi, Wen (2019) An Isotope-Coded Photocleavable Probe for Quantitative Profiling of Protein O-GlcNAcylation. ACS Chemical Biology, 14 (1). pp. 4-10. ISSN 1554-8929. http://resolver.caltech.edu/CaltechAUTHORS:20190109-085256255

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Abstract

O-linked N-acetylglucosamine (O-GlcNAc) is a ubiquitous post-translational modification of proteins and is essential for cell function. Quantifying the dynamics of O-GlcNAcylation in a proteome-wide level is critical for uncovering cellular mechanisms and functional roles of O-GlcNAcylation in cells. Here, we develop an isotope-coded photocleavable probe for profiling protein O-GlcNAcylation dynamics using quantitative mass spectrometry-based proteomics. This probe enables selective tagging and isotopic labeling of O-GlcNAcylated proteins in one step from complex cellular mixtures. We demonstrate the application of the probe to quantitatively profile O-GlcNAcylation sites in 293T cells upon chemical induction of O-GlcNAc levels. We further applied the probe to quantitatively analyze the stoichiometry of O-GlcNAcylation between sorafenib-sensitive and sorafenib-resistant liver cancer cells, which lays the foundation for mechanistic investigation of O-GlcNAcylation in regulating cancer chemoresistance. Thus, this probe provides a powerful tool to profile O-GlcNAcylation dynamics in cells.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1021/acschembio.8b01052DOIArticle
https://pubs.acs.org/doi/suppl/10.1021/acschembio.8b01052PublisherSupporting Information
ORCID:
AuthorORCID
Cai, Li0000-0002-6098-1168
Hsieh-Wilson, Linda C.0000-0001-5661-1714
Yi, Wen0000-0002-4257-3355
Additional Information:© 2019 American Chemical Society. Received: December 4, 2018; Accepted: January 4, 2019; Published: January 8, 2019. This work was supported by the National Science Foundation of China (NSFC, Grant Nos. 91753125, 31322019, 31570804, and 81673387), the National Key Research and Development Program of China (No. 2016YFA0100303), Beijing Nova Program (No. Z161100004916163), and the National Science Foundation of Zhejiang Province (No. LR15C050001). The authors declare no competing financial interest.
Funders:
Funding AgencyGrant Number
National Natural Science Foundation of China91753125
National Natural Science Foundation of China31322019
National Natural Science Foundation of China31570804
National Natural Science Foundation of China81673387
National Key Research and Development Program of China2016YFA0100303
Beijing Nova ProgramZ161100004916163
National Science Foundation of Zhejiang ProvinceLR15C050001
Record Number:CaltechAUTHORS:20190109-085256255
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20190109-085256255
Official Citation:An Isotope-Coded Photocleavable Probe for Quantitative Profiling of Protein O-GlcNAcylation. Jingchao Li, Zhonghua Li, Xiaotao Duan, Ke Qin, Liuyi Dang, Shisheng Sun, Li Cai, Linda C. Hsieh-Wilson, Liming Wu, and Wen Yi. ACS Chemical Biology 2019 14 (1), 4-10. DOI: 10.1021/acschembio.8b01052
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:92154
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:09 Jan 2019 17:05
Last Modified:22 Jan 2019 22:12

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