Protein surface mapping by chemical oxidation: Structural analysis by mass spectrometry

Joshua S. Sharp; Jeffrey M. Becker; Robert L. Hettich

doi:10.1016/S0003-2697(02)00612-7

Protein surface mapping by chemical oxidation: Structural analysis by mass spectrometry

Joshua S. Sharp, Jeffrey M. Becker, Robert L. Hettich

Bioanalytical Mass Spectrometry

Research output: Contribution to journal › Article › peer-review

126 Scopus citations

Abstract

The solvent-accessible surface area of proteins is important in biological function for many reasons, including protein-protein interactions, protein folding, and catalytic sites. Here we present a chemical technique to oxidize amino acid side chains in a model protein, apomyoglobin, and subsequent elucidation of the effect of solvent accessibility on the sites of oxidation. Under conditions of low protein oxidation (zero to three oxygen atoms added per apomyoglobin molecule), we have positively identified five oxidation sites by liquid chromatography-tandem mass spectrometry and high-resolution Fourier transform mass spectrometry. Our results indicate that all oxidized amino acids, with the exception of methionine, have highly solvent-accessible side chains, but the rate of oxidation may not be dictated solely by solvent accessibility and amino acid identity.

Original language	English
Pages (from-to)	216-225
Number of pages	10
Journal	Analytical Biochemistry
Volume	313
Issue number	2
DOIs	https://doi.org/10.1016/S0003-2697(02)00612-7
State	Published - Feb 15 2003

Funding

Research was conducted under the Center for Structural and Molecular Biology Research Program at Oak Ridge National Laboratory (ORNL) and was sponsored by the U.S. Department of Energy, Office of Biological and Environmental Research. The authors are grateful to Nathan VerBerkmoes, Keiji Asano, and Karin Keller for technical assistance with LC-MS/MS experiments. J.S.S. acknowledges financial support from the Graduate School of Genome Science and Technology, University of Tennessee-Oak Ridge National Laboratory. ORNL is operated and managed by the University of Tennessee-Battelle, LLC, for the U.S. Department of Energy under Contract DE-AC05-00OR22725.

Funders	Funder number
Graduate School of Genome Science and Technology
Office of Biological and Environmental Research
University of Tennessee-Battelle	DE-AC05-00OR22725
University of Tennessee-Oak Ridge National Laboratory
U.S. Department of Energy
Oak Ridge National Laboratory

Keywords

Collisional dissociation
Electrospray
Fenton chemistry
Mass spectrometry
Oxidation
Protein derivatization

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10.1016/S0003-2697(02)00612-7

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abstract = "The solvent-accessible surface area of proteins is important in biological function for many reasons, including protein-protein interactions, protein folding, and catalytic sites. Here we present a chemical technique to oxidize amino acid side chains in a model protein, apomyoglobin, and subsequent elucidation of the effect of solvent accessibility on the sites of oxidation. Under conditions of low protein oxidation (zero to three oxygen atoms added per apomyoglobin molecule), we have positively identified five oxidation sites by liquid chromatography-tandem mass spectrometry and high-resolution Fourier transform mass spectrometry. Our results indicate that all oxidized amino acids, with the exception of methionine, have highly solvent-accessible side chains, but the rate of oxidation may not be dictated solely by solvent accessibility and amino acid identity.",

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AU - Becker, Jeffrey M.

AU - Hettich, Robert L.

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N2 - The solvent-accessible surface area of proteins is important in biological function for many reasons, including protein-protein interactions, protein folding, and catalytic sites. Here we present a chemical technique to oxidize amino acid side chains in a model protein, apomyoglobin, and subsequent elucidation of the effect of solvent accessibility on the sites of oxidation. Under conditions of low protein oxidation (zero to three oxygen atoms added per apomyoglobin molecule), we have positively identified five oxidation sites by liquid chromatography-tandem mass spectrometry and high-resolution Fourier transform mass spectrometry. Our results indicate that all oxidized amino acids, with the exception of methionine, have highly solvent-accessible side chains, but the rate of oxidation may not be dictated solely by solvent accessibility and amino acid identity.

AB - The solvent-accessible surface area of proteins is important in biological function for many reasons, including protein-protein interactions, protein folding, and catalytic sites. Here we present a chemical technique to oxidize amino acid side chains in a model protein, apomyoglobin, and subsequent elucidation of the effect of solvent accessibility on the sites of oxidation. Under conditions of low protein oxidation (zero to three oxygen atoms added per apomyoglobin molecule), we have positively identified five oxidation sites by liquid chromatography-tandem mass spectrometry and high-resolution Fourier transform mass spectrometry. Our results indicate that all oxidized amino acids, with the exception of methionine, have highly solvent-accessible side chains, but the rate of oxidation may not be dictated solely by solvent accessibility and amino acid identity.

KW - Collisional dissociation

KW - Electrospray

KW - Fenton chemistry

KW - Mass spectrometry

KW - Oxidation

KW - Protein derivatization

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Protein surface mapping by chemical oxidation: Structural analysis by mass spectrometry

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