Inactivation of rabbit muscle glycogen phosphorylase b by peroxynitrite revisited: Does the nitration of Tyr613 in the allosteric inhibition site control enzymatic function?
| Autor: | Victor S. Sharov, Elena S. Dremina, Todd D. Williams, Nadezhda A. Galeva, Christian Schöneich |
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| Rok vydání: | 2009 |
| Předmět: |
Models
Molecular Protein Conformation Molecular Sequence Data Allosteric regulation Biophysics Biochemistry Article chemistry.chemical_compound Glycogen phosphorylase Protein structure Peroxynitrous Acid Nitration Animals Trypsin Amino Acid Sequence Cysteine Tyrosine Muscle Skeletal Molecular Biology Nitrates Peptide Fragments Kinetics Peroxynitrous acid chemistry Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Glycogen Phosphorylase Muscle Form Rabbits Allosteric Site Peroxynitrite |
| Zdroj: | Archives of Biochemistry and Biophysics. 484:155-166 |
| ISSN: | 0003-9861 |
| DOI: | 10.1016/j.abb.2008.12.012 |
| Popis: | There is increasing evidence that sequence-specific formation of 3-nitrotyrosine (3-NT) may cause functional changes in target proteins. Recently, the nitration of Tyr residues in glycogen phosphorylase b (Ph-b) was implicated in the age-associated decline of protein function [Sharov et al., Exp. Gerontol. 41 (2006) 407–416]; in another report, the nitration of one specific residue, Tyr 613 , located in the allosteric inhibition site was hypothesized as a rationale for peroxynitrite inactivation [Dairou et al., J. Mol. Biol. 372 (2007) 1009–1021]. In this study, we have optimized the analysis of in-gel Ph-b digests by high performance liquid chromatography-electro spray ionization-tandem mass spectrometry, in order to achieve a quantitative analysis of nitration of individual Tyr residues at a high coverage of Tyr-containing sequences (92%). Our data do not confirm the role of Tyr 613 nitration in the control of enzymatic function. Furthermore, we show here that the enzymatic activity of Ph-b does not directly correlate with the protein nitration levels, and that the modification of Cys and, potentially, other amino acid residues can better rationalize Ph-b inactivation by peroxynitrite. |
| Databáze: | OpenAIRE |
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