The long term goals of this research involve modification of the peptidyltransferase (PTC) center in bacterial ribosomes to enable the incorporation of very unusual amino acids not recognized by wild- type ribosomes. In recent years, we have described the selection of modified ribosomes and their use in incorporating D-amino acids, beta amino acids, dipeptides and dipeptidomimetric cassettes into proteins. During the four years of proposed research, the focus of efforts will be on the selection of ribosomes capable of introducing phosphorylated and glycosylated amino acids into specific proteins. The utility of this approach will be explored by analyzing the altered function of proteins into which phosphates and carbohydrates have been introduced, and by expressing a phosphorylated DNA binding protein in E. coli.

Public Health Relevance

The present project is aimed at employing modified bacterial ribosomes to incorporate phosphorylated and glycosylated amino acids into specific proteins. These modified proteins will have their properties altered by the introduction of phosphate and sugar moieties, enabling protein behavior to be better understood by the use of substrates of defined structure in systems that rely on the presence of functional groups usually added posttranslationally. Also proposed is the in cellulo introduction of phosphotyrosine into a DNA binding protein.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Synthetic and Biological Chemistry B Study Section (SBCB)
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Bond, Michelle Rueffer
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Arizona State University-Tempe Campus
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United States
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Chen, Shengxi; Maini, Rumit; Bai, Xiaoguang et al. (2017) Incorporation of Phosphorylated Tyrosine into Proteins: In Vitro Translation and Study of Phosphorylated I?B-? and Its Interaction with NF-?B. J Am Chem Soc 139:14098-14108
Bai, Xiaoguang; Talukder, Poulami; Daskalova, Sasha M et al. (2017) Enhanced Binding Affinity for an i-Motif DNA Substrate Exhibited by a Protein Containing Nucleobase Amino Acids. J Am Chem Soc 139:4611-4614
Chowdhury, Sandipan Roy; Chauhan, Pradeep S; Dedkova, Larisa M et al. (2016) Synthesis and Evaluation of a Library of Fluorescent Dipeptidomimetic Analogues as Substrates for Modified Bacterial Ribosomes. Biochemistry 55:2427-40
Talukder, Poulami; Dedkova, Larisa M; Ellington, Andrew D et al. (2016) Synthesis of alanyl nucleobase amino acids and their incorporation into proteins. Bioorg Med Chem 24:4177-4187
Gerasimova, Yulia V; Yakovchuk, Petro; Dedkova, Larisa M et al. (2015) Expedited quantification of mutant ribosomal RNA by binary deoxyribozyme (BiDz) sensors. RNA 21:1834-43
Chowdhury, Sandipan Roy; Maini, Rumit; Dedkova, Larisa M et al. (2015) Synthesis of fluorescent dipeptidomimetics and their ribosomal incorporation into green fluorescent protein. Bioorg Med Chem Lett 25:4715-8
Maini, Rumit; Dedkova, Larisa M; Paul, Rakesh et al. (2015) Ribosome-Mediated Incorporation of Dipeptides and Dipeptide Analogues into Proteins in Vitro. J Am Chem Soc 137:11206-9
Maini, Rumit; Chowdhury, Sandipan Roy; Dedkova, Larisa M et al. (2015) Protein Synthesis with Ribosomes Selected for the Incorporation of ?-Amino Acids. Biochemistry 54:3694-706