[Influence of the modification of Phe-tRNA synthetase from Escherichia coli by lysine- and arginine-specific reagent on the ionic interactions of the enzyme with tRNA Phe].
The influence of modification of Phe-RSase from E. coli MRE-600 by lysine- and arginine-specific reagent 2,4-pentandione on the Phe-RSase.tRNAPhe interactions was investigated. It was shown that modification of Phe-RSase with 2,4-pentandion leads to a decrease of the aminoacylation rate without any influence on the value of Km for tRNAPhe in this reaction and only a slight increase of the value of Kdiss for Phe-RSase.tRNAPhe complex. The log Km (Km-1)--ionic strength dependence for native enzyme and log Kdiss (K-1diss) for native enzyme and two forms modified on arginine and lysine residues were investigated. Results were interpreted quantitatively by Debye--Huckel approximation for two spherical macroions and by Daune approximation assuming that the region of tRNA implicated in ionic interactions is locally a cylindrical polyelectrolyte. It was shown that there are 2-4 electrostatic contacts in Phe-RSase.tRNAPhe interactions in limits of both approximations; modification of arginine residues in Phe-RSase doesn't change the number of electrostatic contacts, modification of lysine residues leads to an increase in the number of contacts. It was assumed that there are lysine residues in Phe-RSase essential for the tRNAPhe recognition. The possibility of participation of negative amino acid residues in electrostatic interactions with tRNAPhe is not excluded.
D000604:Amino Acyl-tRNA Synthetases / Q000378:metabolism*; D001120:Arginine; D004926:Escherichia coli / Q000201:enzymology*; D007659:Ketones / Q000494:pharmacology*; D007700:Kinetics; D008239:Lysine; D010422:Pentanones / Q000494:pharmacology*; D010649:Phenylalanine; D010652:Phenylalanine-tRNA Ligase / Q000378:metabolism*; D012343:RNA, Transfer; D013329:Structure-Activity Relationship
D007659:Ketones; D010422:Pentanones; D010649:Phenylalanine; D012343:RNA, Transfer; D001120:Arginine; D000604:Amino Acyl-tRNA Synthetases; D010652:Phenylalanine-tRNA Ligase; D008239:Lysine
Vliianie modifikatsii fenilalanil-tRNK-sintetazy po ostatkam lizina i arginina na ionnye vzaimodeĭstviia fermenta s tRNK(phe).