Open AccessGenetically Encoded Protein Tyrosine Nitration in Mammalian Cells
Author(s) -
Jason Porter,
Hyo Sang Jang,
Elise M. Van Fossen,
Duy Nguyen,
Taylor Willi,
Richard B. Cooley,
Ryan A. Mehl
Publication year - 2019
Publication title -
acs chemical biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.899
H-Index - 111
eISSN - 1554-8937
pISSN - 1554-8929
DOI - 10.1021/acschembio.9b00371
Subject(s) - tyrosine , nitration , oxidative stress , biochemistry , nitrotyrosine , biology , oxidative phosphorylation , phosphoproteomics , protein tyrosine phosphatase , chemistry , enzyme , protein phosphorylation , protein kinase a , organic chemistry , nitric oxide synthase
Tyrosine nitration has served as a major biomarker for oxidative stress and is present in high abundance in over 50 disease pathologies in humans. While data mounts on specific disease pathways from specific sites of tyrosine nitration, the role of these modifications is still largely unclear. Strategies for installing site-specific tyrosine nitration in target proteins in eukaryotic cells, through routes not dependent on oxidative stress, would provide a powerful method to address the consequences of tyrosine nitration. Developed here is a Methanosarcina barkeri aminoacyl-tRNA synthetase/tRNA pair that efficiently incorporates nitrotyrosine site-specifically into proteins in mammalian cells. We demonstrate the utility of this approach to produce nitrated proteins identified in disease conditions by producing site-specific nitroTyr-containing manganese superoxide dismutase and 14-3-3 proteins in eukaryotic cells.