Age-associated tyrosine nitration of rat skeletal muscle glycogen phosphorylase b: characterization by HPLC-nanoelectrospray-tandem mass spectrometry

Experimental Gerontology
Victor S SharovChristian Schöneich

Abstract

We identified age-dependent post-translational modifications of skeletal muscle glycogen phosphorylase b (Ph-b), isolated from F1 hybrids of Fisher 344 x Brown Norway rats. Ph-b isolated from 34 months old rats showed a statistically significant decrease in specific activity compared to 6 months old animals: 13.8+/-0.7 vs. 20.6+/-0.8 U mg(-1) protein, respectively. Western blot analysis of the purified Ph-b with anti-3-NT antibodies revealed an age-dependent accumulation of 3-nitrotyrosine (3-NT), quantified by reverse-phase HPLC-UV analysis to increase from 0.05+/-0.03 to 0.34+/-0.11 (mol 3-NT/mol Ph-b) for 6 vs. 34 months old rats, respectively. HPLC-nanoelectrospray ionization-tandem mass spectrometry revealed the accumulation of 3-NT on Tyr113, Tyr161 and Tyr573. While nitration of Tyr113 was detected for both young and old rats, 3-NT at positions 161 and 573 was identified only for Ph-b isolated from 34 months old rats. The sequence of the rat muscle Ph-b was corrected based on our protein sequence mapping and a custom rat PHS2 sequence containing 17 differently located amino acid residues was used instead of the database sequence. The in vitro reaction of peroxynitrite with Ph-b resulted in the nitration of multiple Tyr r...Continue Reading

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Citations

Dec 22, 2010·Free Radical Biology & Medicine·Alexander S BaydenGlen E Kellogg
Jun 4, 2014·Mass Spectrometry Reviews·Woon-Seok YeoKwang Pyo Kim
Jan 27, 2007·Journal of Neurochemistry·Giridharan GokulranganChristian Schöneich
Oct 30, 2013·Mass Spectrometry Reviews·Dimitrios Tsikas, Mark W Duncan
Sep 30, 2006·Experimental Gerontology·Christian Schöneich
Oct 19, 2006·Free Radical Biology & Medicine·Christian Schöneich, Victor S Sharov
Dec 10, 2013·Mass Spectrometry Reviews·Xianquan ZhanDominic M Desiderio
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Jan 31, 2007·Journal of Mass Spectrometry : JMS
Apr 12, 2007·Expert Review of Proteomics·Victor S Sharov, Christian Schöneich
Sep 17, 2010·Journal of Receptor and Signal Transduction Research·Vasily A Yakovlev, Ross B Mikkelsen
Feb 9, 2021·American Journal of Physiology. Endocrinology and Metabolism·Sarah J BlackwoodAbram Katz
Mar 7, 2008·Journal of the American Chemical Society·Heinz NeumannJason W Chin

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