Development of a peptide mapping procedure to identify and quantify methionine oxidation in recombinant human α1-antitrypsin

Steven W. Griffiths; Charles L. Cooney

doi:10.1016/S0021-9673(01)01350-4

Development of a peptide mapping procedure to identify and quantify methionine oxidation in recombinant human α1-antitrypsin

Steven W. Griffiths, Charles L. Cooney

Chemical and Petroleum Engineering

Massachusetts Institute of Technology

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

35 Scopus citations

Abstract

A peptide mapping procedure was developed to identify and quantify methionine oxidation in recombinant human α1-antitrypsin. Due to the protein's complex structural biochemistry, chromatographic analysis of methionine containing digest peptides was a significant challenge. However, by using a combination of mass spectrometry, protein engineering, and high-temperature reversed-phase liquid chromatography, we were able to identify methionine residues that are susceptible to oxidation by hydrogen peroxide, and quantify their reactivity. Our results show that five of the protein's 10 methionine residues are susceptible to oxidation at neutral pH, four of which are localized to the active site region.

Original language	British English
Pages (from-to)	133-143
Number of pages	11
Journal	Journal of Chromatography A
Volume	942
Issue number	1-2
DOIs	https://doi.org/10.1016/S0021-9673(01)01350-4
State	Published - 4 Jan 2002

Keywords

Antitrypsin
Hydrogen peroxide
Methionine
Peptides

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10.1016/S0021-9673(01)01350-4

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abstract = "A peptide mapping procedure was developed to identify and quantify methionine oxidation in recombinant human α1-antitrypsin. Due to the protein's complex structural biochemistry, chromatographic analysis of methionine containing digest peptides was a significant challenge. However, by using a combination of mass spectrometry, protein engineering, and high-temperature reversed-phase liquid chromatography, we were able to identify methionine residues that are susceptible to oxidation by hydrogen peroxide, and quantify their reactivity. Our results show that five of the protein's 10 methionine residues are susceptible to oxidation at neutral pH, four of which are localized to the active site region.",

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AU - Griffiths, Steven W.

AU - Cooney, Charles L.

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N2 - A peptide mapping procedure was developed to identify and quantify methionine oxidation in recombinant human α1-antitrypsin. Due to the protein's complex structural biochemistry, chromatographic analysis of methionine containing digest peptides was a significant challenge. However, by using a combination of mass spectrometry, protein engineering, and high-temperature reversed-phase liquid chromatography, we were able to identify methionine residues that are susceptible to oxidation by hydrogen peroxide, and quantify their reactivity. Our results show that five of the protein's 10 methionine residues are susceptible to oxidation at neutral pH, four of which are localized to the active site region.

AB - A peptide mapping procedure was developed to identify and quantify methionine oxidation in recombinant human α1-antitrypsin. Due to the protein's complex structural biochemistry, chromatographic analysis of methionine containing digest peptides was a significant challenge. However, by using a combination of mass spectrometry, protein engineering, and high-temperature reversed-phase liquid chromatography, we were able to identify methionine residues that are susceptible to oxidation by hydrogen peroxide, and quantify their reactivity. Our results show that five of the protein's 10 methionine residues are susceptible to oxidation at neutral pH, four of which are localized to the active site region.

KW - Antitrypsin

KW - Hydrogen peroxide

KW - Methionine

KW - Peptides

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JO - Journal of Chromatography A

JF - Journal of Chromatography A

IS - 1-2

ER -

Development of a peptide mapping procedure to identify and quantify methionine oxidation in recombinant human α1-antitrypsin

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Keywords

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