Development of a methodology based on metal-catalyzed oxidation reactions to determine the metal binding sites in metalloproteins

Jihyeon Lim; Richard W. Vachet

Development of a methodology based on metal-catalyzed oxidation reactions to determine the metal binding sites in metalloproteins

Jihyeon Lim, Richard W. Vachet

Research output: Contribution to conference › Paper › peer-review

Abstract

The metal binding sites in metalloproteins were determined by development of a methodology based on metal-catalyzed oxidation (MCO) reactions. Azurin was reacted with ascorbate in 50 mM Tris-HCL (Ph 7.4) from time intervals ranging from 30 minutes to 4 hours. The reaction solutions were digested with trypsin and/or chymotrypsin and analyzed for modifications. Oxidation was monitored by removing an aliquot at selected time intervals and analyzing the product by HPLC-ESI-MS using a C ₁₈ reversed-phase columns. In the presence of a catalytic amount of Cu(II), ascorbate mediated modification of the peptide via specific oxidation and/or cleavage reactions was observed.

Original language	English (US)
Pages	317-318
Number of pages	2
State	Published - 2002
Externally published	Yes
Event	Proceedings - 50th ASMS Conference on Mass Spectrometry and Allied Topics - Orlando, FL, United States Duration: Jun 2 2002 → Jun 6 2002

Other

Other	Proceedings - 50th ASMS Conference on Mass Spectrometry and Allied Topics
Country/Territory	United States
City	Orlando, FL
Period	6/2/02 → 6/6/02

ASJC Scopus subject areas

Spectroscopy

Cite this

Development of a methodology based on metal-catalyzed oxidation reactions to determine the metal binding sites in metalloproteins. / Lim, Jihyeon; Vachet, Richard W.
2002. 317-318 Paper presented at Proceedings - 50th ASMS Conference on Mass Spectrometry and Allied Topics, Orlando, FL, United States.

Research output: Contribution to conference › Paper › peer-review

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abstract = "The metal binding sites in metalloproteins were determined by development of a methodology based on metal-catalyzed oxidation (MCO) reactions. Azurin was reacted with ascorbate in 50 mM Tris-HCL (Ph 7.4) from time intervals ranging from 30 minutes to 4 hours. The reaction solutions were digested with trypsin and/or chymotrypsin and analyzed for modifications. Oxidation was monitored by removing an aliquot at selected time intervals and analyzing the product by HPLC-ESI-MS using a C 18 reversed-phase columns. In the presence of a catalytic amount of Cu(II), ascorbate mediated modification of the peptide via specific oxidation and/or cleavage reactions was observed.",

author = "Jihyeon Lim and Vachet, {Richard W.}",

year = "2002",

language = "English (US)",

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note = "Proceedings - 50th ASMS Conference on Mass Spectrometry and Allied Topics ; Conference date: 02-06-2002 Through 06-06-2002",

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AU - Lim, Jihyeon

AU - Vachet, Richard W.

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N2 - The metal binding sites in metalloproteins were determined by development of a methodology based on metal-catalyzed oxidation (MCO) reactions. Azurin was reacted with ascorbate in 50 mM Tris-HCL (Ph 7.4) from time intervals ranging from 30 minutes to 4 hours. The reaction solutions were digested with trypsin and/or chymotrypsin and analyzed for modifications. Oxidation was monitored by removing an aliquot at selected time intervals and analyzing the product by HPLC-ESI-MS using a C 18 reversed-phase columns. In the presence of a catalytic amount of Cu(II), ascorbate mediated modification of the peptide via specific oxidation and/or cleavage reactions was observed.

AB - The metal binding sites in metalloproteins were determined by development of a methodology based on metal-catalyzed oxidation (MCO) reactions. Azurin was reacted with ascorbate in 50 mM Tris-HCL (Ph 7.4) from time intervals ranging from 30 minutes to 4 hours. The reaction solutions were digested with trypsin and/or chymotrypsin and analyzed for modifications. Oxidation was monitored by removing an aliquot at selected time intervals and analyzing the product by HPLC-ESI-MS using a C 18 reversed-phase columns. In the presence of a catalytic amount of Cu(II), ascorbate mediated modification of the peptide via specific oxidation and/or cleavage reactions was observed.

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M3 - Paper

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T2 - Proceedings - 50th ASMS Conference on Mass Spectrometry and Allied Topics

Y2 - 2 June 2002 through 6 June 2002

ER -

Development of a methodology based on metal-catalyzed oxidation reactions to determine the metal binding sites in metalloproteins

Abstract

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ASJC Scopus subject areas

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