Studies on Prolyl-tRNA Synthetase from E. Coli

Studies presented in this dissertation are concerned with both the isolation and characterization of prolyl-tRNA synthetase from E. coli. A detailed analysis of the amino acid binding site was undertaken as were kinetic studies leading to the establishment of the order of addition of substrates and release of products. Evidence that the enzyme possesses different areas for accepting the amino acid and tRNA has been presented; this was demonstrated by the differential affect of temperature on the activity of the enzyme as measured by ATP-PP exchange and esterification reactions. Some of the parameters involved in the interaction of synthetase with tRNA were examined by studying the effect of tRNA and modified tRNA on the protection of the enzyme against cold inactivation. Some of these results have been published previously (1,2,3)