A new type of chemically modified tRNA as a tool for the study of tRNA-aminoacyl-tRNA synthetase interaction.
tRNA(Phe) in which the adenine and cytosine rings in the aminoacyl arm and in the anticodon loop were converted to alkylating derivatives by mild treatment with methyl chlorotetrolate was used to study the tRNA(Phe)-yeast phenylalanyl-tRNA(Phe) synthetase interaction. At neutral pH, modified tRNA inhibited the enzyme competitively. At pH 9 this binding is accompanied by irreversible inactivation of the enzyme due to alkylation of the alpha subunit of the synthetase. Such a derivatization of tRNA could probably be used to investigate the interaction of other tRNAs with their cognate synthetases.
Roques
P
P
Laboratoire de Biochimie Cellulaire, Collège de France, Paris.
Thomé
F
F
Dubord
C
C
Olomucki
M
M
D000478:Alkylation; D000480:Alkynes; D000604:Amino Acyl-tRNA Synthetases / Q000037:antagonists & inhibitors*; D001667:Binding, Competitive; D055598:Chemical Phenomena; D002621:Chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D015394:Molecular Structure; D010652:Phenylalanine-tRNA Ligase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D012345:RNA, Transfer, Amino Acid-Specific / Q000494:pharmacology*; D012360:RNA, Transfer, Phe / Q000378:metabolism / Q000494:pharmacology*; D012441:Saccharomyces cerevisiae / Q000201:enzymology
D000480:Alkynes; D012345:RNA, Transfer, Amino Acid-Specific; D012360:RNA, Transfer, Phe; C051334:methyl chlorotetrolate; D000604:Amino Acyl-tRNA Synthetases; D010652:Phenylalanine-tRNA Ligase