Evaluation of D10-Leu Metabolic Labeling Coupled with MALDI− MS

Synopsis. Response of the S. cerevisiae proteome to H2O2 was probed by D10-Leu metabolic-labeling and isotope-ratio quantitation. Exponential-phase ce...
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Evaluation of D10-Leu Metabolic Labeling Coupled with MALDI-MS Analysis in Studying the Response of the Yeast Proteome to H2O2 Challenge Heng Jiang and Ann M. English* Centre for Biological Applications of Mass Spectrometry, Department of Chemistry and Biochemistry, Concordia University, Montreal, Quebec, Canada H4B 1R6 Received January 19, 2006

An efficient D10-Leu metabolic-labeling method combined with isotope-ratio quantitation by MALDITOF MS was used to probe the response of the yeast proteome to H2O2. Control cultures correct for effects not associated with H2O2 challenge. A stress-response index to H2O2 (SRIH2O2) is defined, and values are reported for seven proteins at 45-225 min following exposure to 0.4 mM H2O2. The time course of protein accumulation in unstressed cells following the H10- to D10-SCD switch suggests that proteome responses at 1 ( 1 (