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PROTEOMIC ANALYSIS OPTIMIZATION: SELECTIVE PROTEIN SAMPLE ON-COLUMN RETENTION IN REVERSE-PHASE LIQUID CHROMATOGRAPHY
Citation:
WINNIK, W. M. AND P. A. ORTIZ. PROTEOMIC ANALYSIS OPTIMIZATION: SELECTIVE PROTEIN SAMPLE ON-COLUMN RETENTION IN REVERSE-PHASE LIQUID CHROMATOGRAPHY. Journal of Chromatography B. Elsevier Science Ltd, New York, NY, 233(1):478-486, (2008).
Impact/Purpose:
To be able to identify, on a proteomic level, cytochromes P450 (CYP) and UDP-glucuronosyltransferases (UGT) in mouse liver microsomes for the conazole exposure study. The new enrichment method was necessary because the available proteomic methods did not provide sufficient sensitivity and selectivity of protein analysis.
Description:
Why work was done?
To be able to identify, on a proteomic level, cytochromes P450 (CYP) and UDP-glucuronosyltransferases (UGT) in mouse liver microsomes for the conazole exposure study IRP # NHEERL-ECD-SCN-CZ-2002-01-R1_Addendum 1. The new enrichment method was necessary because the available proteomic methods did not provide sufficient sensitivity and selectivity of protein analysis.
What is the impact to the field and the agency?
Cytochromes P450 (CYP) and UDP-glucuronosyltransferases (UGT) are phase I and phase II enzymes, respectively, representing diverse families of membrane proteins. CYP enzymes are instrumental in the oxidative metabolism of endogenous and xenobiotic compounds. The UGTs participate in the conjugation of hydrophobic compounds with glucuronic acid to increase their water solubility and facilitate excretion. Induction of cytochrome P450 genes and their mutations and modifications are responsible for a variety of human diseases. For example, induction of P450s by environmental compounds might be a cancer risk factor because CYPs can convert pro-carcinogens to carcinogens. Susceptibility to environmental xenobiotics and toxicity depend on the expression level and efficiency of these enzymes. Because of these health implications, the identification and quantification of the CYP and UGT proteins on the genomic and proteomic levels are used in studies of toxic effects of chemicals in the environment, drug interactions and metabolism, and in clinical and cancer research.
The method will be applied in a CYP and UGT study of mouse microsomal protein fractions obtained from the animals exposed to conazole fungicides. The correlation between these proteomic and genomic results is expected to increase the knowledge base for the mode of action of this class of compounds.
In general, the expression of CYPs and UGTs are of prime interest to the scientists working in multiple research areas, such as drug metabolism and interactions, environmental xenobiotic toxicity, and carcinogenesis. The new analytical method is anticipated to impact such proteomic-based research projects.