Detailed map of oxidative post-translational modifications of human p21ras using Fourier transform mass spectrometry.

Detailed map of oxidative post-translational modifications of human p21ras using Fourier transform mass spectrometry. Research Abstract Details 

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Detailed map of oxidative post-translational modifications of human p21ras using Fourier transform mass spectrometry. Abstract Text:

Cheng Zhao,Mahadevan Sethuraman,Nicolas Clavreul,Parminder Kaur,Richard A Cohen,Peter B O'Connor,

P21ras, the translation product of the most commonly mutated oncogene, is a small guanine nucleotide exchange protein. Oxidant-induced post-translational modifications of p21ras including S-nitrosation and S-glutathiolation have been demonstrated to modulate its activity. Structural characterization of this protein is critical to further understanding of the biological functions of p21ras. In this study, high-resolution and high mass accuracy Fourier transform mass spectrometry was utilized to map, in detail, the post-translational modifications of p21ras (H-ras) exposed to oxidants by combining bottom-up and top-down techniques. For peroxynitrite-treated p21ras, five oxidized methionines, five nitrated tyrosines, and at least two oxidized cysteines (including C118) were identified by "bottom-up" analysis, and the major oxidative modification of C118, Cys118-SO3H, was confirmed by several tandem mass spectrometry experiments. Additionally, "top-down" analysis was conducted on p21ras S-glutathiolated by oxidized glutathione and identified C118 as the major site of glutathiolation among the four surface cysteines. The present study provides a paradigm for an effective and efficient method not only for mapping post-translational modifications of proteins but also for predicting the relative selectivity and specificity of oxidative post-translational modifications, especially using top-down analysis.

Detailed map of oxidative post-translational modifications of human p21ras using Fourier transform mass spectrometry. Publishing Authors By Initials

C Zhao,M Sethuraman,N Clavreul,P Kaur,RA Cohen,PB O'Connor,

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Detailed map of oxidative post-translational modifications of human p21ras using Fourier transform mass spectrometry. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Analytical chemistry

VOLUME: 78

Page Numbers: 5134-42

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ISSN: 0003-2700

DAY: 15

MONTH: Jul

YEAR: 2006

Detailed map of oxidative post-translational modifications of human p21ras using Fourier transform mass spectrometry. Information

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LANGUAGE: eng

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AFFILIATION: Mass Spectrometry Resource, Department of Biochemistry, Cardiovascular Proteomics Center, and Vascular Biology Unit, Boston University School of Medicine, Boston, Massachusetts 02118, USA.

Country: United States

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MEDLINETA: Anal Chem

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