Reaction of tryptophanyl-tRNA synthetase from beef pancreas with periodate-oxidized ATP.
Tryptophanyl-tRNA synthetase from beef pancreas reacts with periodate-oxidized ATP according to biphasic kinetics. A rapid phase involves two groups of the protein, presumably lysine side-chains. The slow phase corresponds to the reaction of a larger number of groups. The time-course of the partial losses of the ATP-PPi isotopic exchange and of the aminoacylation activities of the enzyme follow the labelling of the two fast-reacting groups. However, the ability of the enzyme to form a bis(tryptophanyladenylate)-enzyme complex is not lost after reaction of these two groups with the reagent. The affinity for ATP is also unaffected by this initial labelling of the protein, as seen from the Km values of this substrate in the ATP-PPi isotopic exchange reaction. These data suggest that, in this fast initial reaction, oxidized ATP reacts neither with specific ATP-binding groups of the enzyme nor with any major catalytic residue of the tryptophan-activation site. In contrast with this first step, the further slow labelling of lysine residues leads to a disappearance of the aminoacylation ability of the enzyme, while it does not further affect the ATP-PPi exchange activity. The behaviour of beef tryptophanyl-tRNA synthetase during derivatization with oxidized ATP is therefore at variance with that which has been described for the homologous E. coli enzyme.
Fournier
M
M
Institut de Biochimie Cellulaire et Neurochimie, Centre National de la Recherche Scientifique, Bordeaux, France.
Labouesse
B
B
Labouesse
J
J
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000378:metabolism; D000604:Amino Acyl-tRNA Synthetases / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D011756:Diphosphates / Q000378:metabolism; D007700:Kinetics; D010179:Pancreas / Q000201:enzymology*; D012364:RNA, Transfer, Trp / Q000378:metabolism; D014369:Tryptophan-tRNA Ligase / Q000378:metabolism*
D011756:Diphosphates; D012364:RNA, Transfer, Trp; C004282:periodate-oxidized adenosine 5'-triphosphate; D000255:Adenosine Triphosphate; D000604:Amino Acyl-tRNA Synthetases; D014369:Tryptophan-tRNA Ligase