ARGINYL-tRNA SYNTHETASE FROM Escherichia coli AFFINITY LABELING WITH 3'-OXIDIZED tRNA~(Arg)

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The covalent modification of E. coli arginyl-tRNA synthetase by the 2’,3’-dialdehydederivative of tRNA~(Arg) (tRNA_(ox)~(Arg)) resulted in the complete inactivation of the ATP-PPi ex-change and aminoacylation activities of the enzyme. Sodium dodecyl sulfate polyacrylamide gelelectrophoresis of the ArgRS-tRNA_(ox)~(Arg) covalent complexes indicated that two bands simulta-neously appeared on the gel parallel with inactivation corresponding to different higher mo-lecular weights. This result was different from that of the other aminoacyl-tRNA synthetaselabeling systems as previously reported. Upon the ribonuclease treatment of the modifiedArgRS, less than 15% of both the initial ATP-PPi exchange and aminocylation activities wererecovered. During the whole process of labeling and RNase treatment, the two activities ofthe enzyme were closely associated. The covalent modification of E. coli arginyl-tRNA synthetase by the 2 ’, 3’-dialdehydederivative of tRNA ~ (Arg) (tRNA_ (Arg) ~ (Arg)) in the complete inactivation of the ATP-PPi ex-change and aminoacylation activities of the enzyme. Sodium dodecyl sulfate polyacrylamide gelelectrophoresis of the ArgRS-tRNA_ (ox) ~ (Arg) covalent complexes indicated that two bands simulta-nerous appeared on the gel parallel with inactivation corresponding to different higher mo-lecular weights. This result Upon the ribonuclease treatment of the modified ArgRS, less than 15% of both the initial ATP-PPi exchange and aminocylation activities wererecovered. During the whole process of labeling and RNase treatment the two activities of the enzyme were closely associated.
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