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 conferencePaper

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)
Pages317-318
Number of pages2
StatePublished - Dec 1 2002
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

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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. 317-318. Paper presented at Proceedings - 50th ASMS Conference on Mass Spectrometry and Allied Topics, Orlando, FL, United States.