This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.The proposed experiments investigate the structural basis of enzyme-RNA recognition and catalysis. Experiments will encompass Xray crystallographic studies of novel liganded states and of modified complexes for both the glutaminyl-tRNA synthetase and cysteinyl-tRNA synthetase complexes from E. coli. Emphasis will be placed on elucidating the stereochemical pathways of cleavage and on Xray studies that will support parallel protein engineering work to alter amino acid specificity, thus offering the potential to expand the genetic code to include incorporation of nonstandard amino acids. Other experiments will address the complex roles of indirect readout and induced-fit conformational change in conferring the very high selectivities for transfer RNA and for amino acid that are characteristic of tRNA synthetases.
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