Funds are requested for the acquisition of a CEM Liberty automated microwave-assisted solid phase peptide synthesizer and associated accessories so that we may employ this advanced technology in a variety of synthesis, proteomic, and glycoproteomic applications, with an emphasis on glycobiology. This will enhance the research efforts of seven currently NIH funded investigators, and a new investigator with a pending NIH application, one with NSF funding and a newly arrived Emminent Scholar with a pending DOE application. Protein glycosylation is the most prevalent and complex mode of protein post-translational processing, being an important factor in health and disease, but studying glycoproteins is often limited by their molecular weight and glycan microheterogeneity. Synthetic glycopeptides provide powerful alternative agents for investigating many aspects of glycoproteins. The additional manipulations associated with glycopeptide synthesis have severely limited their availability relative to peptides, a factor that has been a major limitation is studying properties of glycoproteins. We have been working with more laborious conventional capabilities in this area, and a key factor in this request is the demonstration that the newly developed microwave-assisted methodology has proven particularly beneficial for accelerating glycopeptide synthesis. Microwave-assisted approaches have also proven beneficial in proteolytic enzyme digestions and N-glycan release, addressing the needs of several of the participating investigators in porteomics and glycoproteiomcs, and will therefore help identify biologically important aspects of glycorpoteins and identify relevant glycopeptides to be synthesized for further study. This instrumentation would be located in the Complex Carbohydrate Research Center at the University of Georgia which is a preeminent center in glycobiology research, and provides a unique environment encompassing chemistry, biochemistry, molecular biology and cell biology expertise for optimally exploiting the technology in the rapidly developing field of glycobiology.
The enhanced microwave-assisted technologies in synthesis and proteomics will be employed to advance current research projects primarily focusing on investigating carbohydrate modified proteins. Protein glycosylation is a very important component of post-translational modification with impacts in health and disease, and this instrumentation will facilitate preparation of glycoprotein fragments and analysis of glycoproteins of biomedical significance.
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|Prestegard, James H; Sahu, Sarata C; Nkari, Wendy K et al. (2013) Chemical shift prediction for denatured proteins. J Biomol NMR 55:201-9|