Dataset Information

An azidoribose probe to track ketoamine adducts in histone ribose glycation (293T nuclei - LFQ)

ABSTRACT: Reactive cellular metabolites can modify macromolecules and form adducts known as non-enzymatic covalent modifications (NECMs). Dissecting 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. Directly tracking the formation of modifications on key targets to uncover their underlying physiological importance requires specific chemical tools. Here we present the novel chemoenzymatic syntheses of an active azido-modified ribose analog, 5-azidoribose (5-AR), as well as an inactive control derivative, 1-azidoribose (1-AR) and their application towards understanding protein ribose-glycation in vitro and in cellulo. With these new probes we found that, similar to 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 highresolution 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 chemical tool to monitor FN3K activity. Altogether, we demonstrate this probe’s utilities to uncover ribose-glycation and deglycation events as well as tracking FN3K activity towards establishing its potential as a new cancer vulnerability.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell

DISEASE(S): Disease Free

SUBMITTER: Igor Maksimovic

LAB HEAD: Yael David

PROVIDER: PXD019204 | Pride | 2020-05-15

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	MS205917QEHF_igor_david_1AR_01.raw	Raw
	MS205917QEHF_igor_david_1AR_02_20191017140015.raw	Raw
	MS205917QEHF_igor_david_1AR_03.raw	Raw
	MS205917QEHF_igor_david_5AR_RIPA_01.raw	Raw
	MS205917QEHF_igor_david_5AR_RIPA_02.raw	Raw

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Publications

An Azidoribose Probe to Track Ketoamine Adducts in Histone Ribose Glycation.

Maksimovic Igor I Zheng Qingfei Q Trujillo Marissa N MN Galligan James J JJ David Yael Y

Journal of the American Chemical Society 20200522 22

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 ...[more]

PMID: 32390412

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