Fluorescence-monitored conformational change on the 3'-end of tRNA upon aminoacylation.
Fluorescent tRNAs species with formycine in the 3'-terminal position (tRNA-CCF) were derived from Escherichia coli tRNA(Val). Thermus thermophilus tRNA(Aap) and Thermus thermophilus tRNA(Phe). The fluorescence of formycine was used to monitor the conformational changes at the 3'-terminus of tRNA caused by aminoacylation and hydrolysis of aminoacyl residue from aminoacyl-tRNAs. An increase of about 15% in the fluorescence intensity was observed after aminoacylation of the three tRNA-CCF. This change in fluorescence amplitude that is reversed by hydrolysis of the aminoacyl residue, does not depend on the structure of the amino acid or tRNA sequence. A local conformational change at the 3'-terminal formycine probably involving a partial destacking of the base moiety in the ACCF end takes place as a consequence of aminoacylation. A structural change at the 3'-terminus of tRNA induced by attachment and detachment of the acyl residue may be important in controlling the substrate/product relationship in reactions in which tRNA participates during protein biosynthesis.
Schlosser
A
A
Laboratorium für Biochemie Universität Bayreuth, Germany.
Nawrot
B
B
Grillenbeck
N
N
Sprinzl
M
M
D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D005573:Formycins / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D009690:Nucleic Acid Conformation; D012346:RNA, Transfer, Amino Acyl / Q000737:chemistry*; D012349:RNA, Transfer, Asp / Q000737:chemistry; D012360:RNA, Transfer, Phe / Q000737:chemistry; D012366:RNA, Transfer, Val / Q000737:chemistry; D012265:Ribonucleotides / Q000737:chemistry; D016963:Thermus thermophilus / Q000737:chemistry
D005456:Fluorescent Dyes; D005573:Formycins; D012346:RNA, Transfer, Amino Acyl; D012349:RNA, Transfer, Asp; D012360:RNA, Transfer, Phe; D012366:RNA, Transfer, Val; D012265:Ribonucleotides; C032552:formycin triphosphate