Proteomics is the study of protein expression at a global level for a specific cell type, tissue or organ. A primary objective of Proteomics is the identification of differentially expressed proteins after experimental treatment, environmental contaminant exposure or in human disease. Understanding global changes in gene expression at the protein level will be a useful tool for experimental biology, can implicate new targets or pathways for therapeutics, and will provide new markers for environmental toxicology. Proteomics is accomplished by combining protein separation and identification technologies in a sequential and automated fashion. We are utilizing the high resolution capabilities of two-dimensional gel electrophoresis for protein separation and mass spectrometry technologies for protein identification. The Regulatory Proteins (RP) Group in LMC and Mass Spectrometry (MS) Group in LSB have devoted personnel and resources to form a Proteomics working group that operates within a cross-laboratory network termed the NIEHS Gene Expression Center. The RP and MS groups have established a work flow for Proteomics that involves 2D PAGE protein separation of any tissue sample, detection, imaging for differential protein expression, protein excision and digestion in 96 well format, MS analysis and database searching for protein profiling of the sample. MALDI/MS is used for protein fingerprint screening from public databases. Tandem mass spectrometry (MS/MS) is selectively used for protein sequencing and microcharacterization studies. The Proteomics working group is completing proof-of-principle studies and is engaged in collaborative studies with several DIR research groups at NIEHS.
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