Publication Information
Doron et al., 2021
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Abstract
Free Radic Biol Med. 2021 Nov 20;176:366-377. doi:
10.1016/j.freeradbiomed.2021.10.001. Epub 2021 Oct 5.
SPEAR: A proteomics approach for simultaneous protein expression and redox
analysis.
Doron S(1), Lampl N(1), Savidor A(2), Katina C(2), Gabashvili A(2), Levin Y(3),
Rosenwasser S(4).
Author information:
(1)The Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture,
The Hebrew University of Jerusalem, Rehovot 7610000, Israel.
(2)de Botton Institute for Protein Profiling, The Nancy and Stephen Grand Israel
National Center for Personalized Medicine, Weizmann Institute of Science,
Rehovot, Israel.
(3)de Botton Institute for Protein Profiling, The Nancy and Stephen Grand Israel
National Center for Personalized Medicine, Weizmann Institute of Science,
Rehovot, Israel. Electronic address: yishai.levin@weizmann.ac.il.
(4)The Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture,
The Hebrew University of Jerusalem, Rehovot 7610000, Israel. Electronic address:
shilo.rosenwaser@mail.huji.ac.il.
Oxidation and reduction of protein cysteinyl thiols serve as molecular switches,
which is considered the most central mechanism for redox regulation of
biological processes, altering protein structure, biochemical activity,
subcellular localization, and binding affinity. Redox proteomics allows global
identification of redox-modified cysteine (Cys) sites and quantification of
their reversible oxidation/reduction responses, serving as a
hypothesis-generating platform to stimulate redox biology mechanistic research.
Here, we developed Simultaneous Protein Expression and Redox (SPEAR) analysis, a
new redox-proteomics approach based on differential labeling of reversibly
oxidized and reduced cysteines with light and heavy isotopic forms of
commercially available isotopically-labeled N-ethylmaleimide (NEM). The
presented method does not require enrichment for labeled peptides, thus enabling
simultaneous quantification of Cys reversible oxidation state and protein
abundance. Using SPEAR, we were able to quantify the in-vivo reversible
oxidation state of thousands of cysteines across the Arabidopsis proteome under
steady-state and oxidative stress conditions. Functional assignment of the
identified redox-sensitive proteins demonstrated the widespread effect of
oxidative conditions on various cellular functions and highlighted the
enrichment of chloroplastic proteins. SPEAR provides a simple, straightforward,
and cost-effective means of studying redox proteome dynamics. The presented data
provide a global quantitative view of the reversible oxidation of well-known
redox-regulated active sites and many novel redox-sensitive sites whose role in
plant acclimation to stress conditions remains to be further explored.
Copyright © 2021 Elsevier Inc. All rights reserved.
DOI: 10.1016/j.freeradbiomed.2021.10.001
PMID: 34619326 [Indexed for MEDLINE]
