An azidoribose probe to track ketoamine adducts in histone ribose glycation Journal Article


Authors: Maksimovic, I.; Zheng, Q.; Trujillo, M. N.; Galligan, J. J.; David, Y.
Article Title: An azidoribose probe to track ketoamine adducts in histone ribose glycation
Abstract: Reactive cellular metabolites can modify macromolecules and form adducts known as nonenzymatic covalent modifications (NECMs). The dissection of the mechanisms, regulation, and consequences of NECMs, such as glycation, has been challenging due to the complex and often ambiguous nature of the adducts formed. Specific chemical tools are required to directly track the formation of these modifications on key targets in order to uncover their underlying physiological importance. Here, we present the novel chemoenzymatic synthesis of an active azido-modified ribose analog, 5-azidoribose (5-AR), as well as the synthesis of an inactive control derivative, 1-azidoribose (1-AR), and their application toward understanding protein ribose-glycation in vitro and in cellulo. With these new probes we found that, similar to methylglyoxal (MGO) glycation, ribose glycation specifically accumulates on histones. In addition to fluorescent labeling, we demonstrate the utility of the probe in enriching modified targets, which were identified by label-free quantitative proteomics and high-resolution MS/MS workflows. Finally, we establish that the known oncoprotein and hexose deglycase, fructosamine 3-kinase (FN3K), recognizes and facilitates the removal of 5-AR glycation adducts in live cells, supporting the dynamic regulation of ribose glycation as well as validating the probe as a new platform to monitor FN3K activity. Altogether, we demonstrate this probe's utilities to uncover ribose-glycation and deglycation events as well as track FN3K activity toward establishing its potential as a new cancer vulnerability. Copyright © 2020 American Chemical Society.
Keywords: proteins; proteomics; biosynthesis; glycosylation; probes; high resolution; chemical modification; covalent modifications; metabolites; quantitative proteomics; dynamic regulation; cellular metabolites; chemo-enzymatic synthesis; control derivatives; fluorescent labeling
Journal Title: Journal of the American Chemical Society
Volume: 142
Issue: 22
ISSN: 0002-7863
Publisher: American Chemical Society  
Date Published: 2020-06-03
Start Page: 9999
End Page: 10007
Language: English
DOI: 10.1021/jacs.0c01325
PUBMED: 32390412
PROVIDER: scopus
DOI/URL:
Notes: Article -- Export Date: 1 July 2020 -- Source: Scopus
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MSK Authors
  1. Qingfei Zheng
    8 Zheng