ANSID: A Solid-Phase Proteomic Approach for Identification and Relative Quantification of Aromatic Nitration Sites

Frontiers in Chemistry
Tal NurielSteven S Gross

Abstract

Nitration of tyrosine and other aromatic amino acid residues in proteins occurs in the setting of inflammatory, neurodegenerative, and cardiovascular diseases-importantly, this modification has been implicated in the pathogenesis of diverse diseases and the physiological process of aging. To understand the biological consequences of aromatic nitration in both health and disease, it is critical to molecularly identify the proteins that undergo nitration, specify their cognate modification sites and quantify their extent of nitration. To date, unbiased identification of nitrated proteins has often involved painstaking 2D-gel electrophoresis followed by Western Blotting with an anti-nitrotyrosine antibody for detection. Apart from being relatively slow and laborious, this method suffers from limited coverage, the potential for false-positive identifications, and failure to reveal specific amino acid modification sites. To overcome these shortcomings, we have developed a solid-phase, chemical-capture approach for unbiased and high-throughput discovery of nitrotyrosine and nitrotryptophan sites in proteins. Utilizing this method, we have successfully identified several endogenously nitrated proteins in rat brain and a total of 244 n...Continue Reading

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Citations

Jul 11, 2018·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Md Mehedi HasanGuo Dianjing
Jun 27, 2019·Chemistry : a European Journal·Yuxin LiuJianguo Fang
Feb 6, 2018·Chemical Reviews·Gerardo Ferrer-SuetaRafael Radi
May 24, 2019·ACS Chemical Biology·Joseph J PorterRyan A Mehl

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Methods Mentioned

BETA
electrophoresis
protein assay
chip
acetylation
affinity capture
sample pooling

Software Mentioned

Mass Hunter Qualitative Analysis
Spectrum Mill
ANSID

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