Fluorescence resonance energy transfer analysis of ribozyme kinetics reveals the mode of action of a facilitator oligonucleotide.

摘要: A defining characteristic of catalysts is the rate at which they can process multiple copies substrate. In case synthetic hammerhead ribozymes that cleave an RNA sequence, binding ribozyme to substrate and products through base-paired duplexes. The kinetics formation dissociation these duplexes determine turnover ribozyme. We have followed processes in real time by using fluorescent labels interact fluorescence resonance energy transfer (FRET). This approach has been used identify rate-limiting steps for a particular reveal how was improved facilitator oligonucleotide. It found ribozyme-substrate complex faster than cleavage products. Hence, undergo cleavage, most molecules must with more once. presence oligonucleotide, stabilized so dissociation. Under circumstances, becomes likely outcome following