Single-molecule, real-time measurement of enzyme kinetics by alternating-laser excitation fluorescence resonance energy transfer.
Using a single-molecule fluorescence method uniquely suitable for binding assay, alternating-laser excitation fluorescence resonance energy transfer (ALEX-FRET), we accurately measured the cleavage rate of 8-17 deoxyribozyme, an RNA-cleaving enzyme, at the single-molecule level in real time with a minimum consumption of samples, i.e., at least three orders of magnitude smaller than used in the conventional ensemble FRET method.
Lee
Nam Ki
NK
Department of Biophysics and Chemical Biology, Seoul National University, Seoul 151-747, Korea.
Koh
Hye Ran
HR
Han
Kyu Young
KY
Lee
Jihyun
J
Kim
Seong Keun
SK
D021881:DNA, Catalytic / Q000737:chemistry* / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D007700:Kinetics; D007834:Lasers; D012313:RNA / Q000737:chemistry / Q000378:metabolism
D021881:DNA, Catalytic; D012313:RNA