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: Chapter in Book/Report/Conference proceedingConference contribution

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 languageEnglish (US)
Title of host publicationProceedings 50th ASMS Conference on Mass Spectrometry and Allied Topics
Pages317-318
Number of pages2
StatePublished - 2002
Externally publishedYes
EventProceedings - 50th ASMS Conference on Mass Spectrometry and Allied Topics - Orlando, FL, United States
Duration: Jun 2 2002Jun 6 2002

Other

OtherProceedings - 50th ASMS Conference on Mass Spectrometry and Allied Topics
CountryUnited States
CityOrlando, FL
Period6/2/026/6/02

Fingerprint

Metalloproteins
Metals
Binding Sites
Oxidation
Azurin
Chymotrypsin
Trypsin
Peptides

ASJC Scopus subject areas

  • Spectroscopy

Cite this

Lim, J., & Vachet, R. W. (2002). Development of a methodology based on metal-catalyzed oxidation reactions to determine the metal binding sites in metalloproteins. In Proceedings 50th ASMS Conference on Mass Spectrometry and Allied Topics (pp. 317-318)

Development of a methodology based on metal-catalyzed oxidation reactions to determine the metal binding sites in metalloproteins. / Lim, Jihyeon; Vachet, Richard W.

Proceedings 50th ASMS Conference on Mass Spectrometry and Allied Topics. 2002. p. 317-318.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Lim, J & Vachet, RW 2002, Development of a methodology based on metal-catalyzed oxidation reactions to determine the metal binding sites in metalloproteins. in Proceedings 50th ASMS Conference on Mass Spectrometry and Allied Topics. pp. 317-318, Proceedings - 50th ASMS Conference on Mass Spectrometry and Allied Topics, Orlando, FL, United States, 6/2/02.
Lim J, Vachet RW. Development of a methodology based on metal-catalyzed oxidation reactions to determine the metal binding sites in metalloproteins. In Proceedings 50th ASMS Conference on Mass Spectrometry and Allied Topics. 2002. p. 317-318
Lim, Jihyeon ; Vachet, Richard W. / Development of a methodology based on metal-catalyzed oxidation reactions to determine the metal binding sites in metalloproteins. Proceedings 50th ASMS Conference on Mass Spectrometry and Allied Topics. 2002. pp. 317-318
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