Using Mass Spectrometry to Assist in Identifying Genes for Enzymes of Known Activity, Unknown OpenReadingFrame and Small Abundance
EMSL Project ID
3683
Abstract
We have reported the identification of the Open Reading Frame (ORF) that expresses a NADPH-dependent methylglyoxal reducing enzyme based on the partial purification of this activity through 3 simple purification steps and the correlation of its specific activity and the relative increase in the intensity of Coomassie Blue stained bands on an SDS/PAGE gel. We propose substituting the relative intensity of mass spectrometry signals at each stage of partial purification for the intensity of bands on a gel. This should allow us to apply this approach to gene products that are expressed at modest to slight abundance. We hope to verify the plausibility of this approach by applying it first to a well understood system.
Project Details
Project type
Exploratory Research
Start Date
2003-10-10
End Date
2005-10-26
Status
Closed
Released Data Link
Team
Principal Investigator