Using fluorescence resonance energy transfer (FRET) for measuring 2-5A analogues ability to activate RNase L.
The development of a method for measuring the ability of 2-5A analogues to activate the cleavage of an oligoribonucleotide substrate by RNase L is described. This method is based on fluorescence resonance energy transfer. The method is easily performed with 96-well plates, allowing for quantitative high-throughput analyses of 2-5A analogues under different reaction conditions.
Cramer
H
H
Section of Biomedical Chemistry, NIDDK, NIH, Bethesda, MD 20892, USA.
Geselowitz
D A
DA
Torrence
P F
PF
D000227:Adenine Nucleotides / Q000737:chemistry* / Q000494:pharmacology; D004722:Endoribonucleases; D004735:Energy Transfer; D004789:Enzyme Activation; D006801:Humans; D007700:Kinetics; D016376:Oligonucleotides, Antisense / Q000737:chemistry; D009843:Oligoribonucleotides / Q000737:chemistry* / Q000494:pharmacology; D013050:Spectrometry, Fluorescence
D000227:Adenine Nucleotides; D016376:Oligonucleotides, Antisense; D009843:Oligoribonucleotides; C023505:2',5'-oligoadenylate; D004722:Endoribonucleases; C031956:2-5A-dependent ribonuclease