The Kresge Center Proteomics Facility (KCPF), which is directed by the PI of this grant, is an NIEHS and NSF sponsored resource at the Harvard School of Public Health (HSPH) that offers an infrastructure for protein analysis at a system-wide level (= """"""""proteomics""""""""). KCPF offers automated proteomics processes based on 2-dimensional gel electrophoresis (2DGE) and protein identification by liquid chromatography and tandem mass spectrometry (LC/MS/MS). In this grant application funds are requested for the acquisition of a combined robotic spot picker and liquid handler, which will be integrated in the existing chain of automated processes that deliver high quality protein analysis data. KCPF offers two major applications for proteomics research that will substantially benefit from the requested equipment: 1. Protein expression profiling, and 2. Identification of protein assemblies purified from complex samples. The research activities resulting from funding of this proposal make extensive use of these applications in order to answer major questions in the basic biology of disease. The projects use samples derived from organisms across the evolutionary ladder, including viruses, bacteria, yeast, parasites, transgenic mice, mammalian tissue culture cells, and human organs. The scope of research activities is wide-ranging, and includes large scale analysis of the ubiquitin-dependent proteolysis system, analyzing determinants of cellular responses to radicals and oxidative stress, identifying proteins involved in placental dysfunction in preterm deliveries, determining factors controlling membrane transport of metals and peptide growth factors, analyzing signaling and transcriptional activation pathways in cell lineage determination, determining protein interactions involved in establishing cell polarity, among many others. Ongoing projects in these areas have revealed an acute need for the capability to rapidly process an extensive number of protein spots identified on 2D gels. This includes spot excision from gels, extraction of the proteins from gel plugs, enzymatic digestions, and purification of the peptides for MS analysis. The recently developed spot picking and liquid handling device requested in this grant application combines all of these capabilities in a single unit.
|Jones, Matthew R; Quinton, Lee J; Blahna, Matthew T et al. (2009) Zcchc11-dependent uridylation of microRNA directs cytokine expression. Nat Cell Biol 11:1157-63|