Dynamic Disorder in Single-Enzyme Experiments: Facts and Artifacts
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- 作者：Tatyana G. Terentyeva ; Hans Engelkamp ; Alan E. Rowan ; Tamiki Komatsuzaki ; Johan Hofkens ; Chun-Biu Li ; Kerstin Blank
- 刊名：ACS Nano
- 出版年：2012
- 出版时间：January 24, 2012
- 年：2012
- 卷：6
- 期：1
- 页码：346-354
- 全文大小：1412K
- 年卷期：v.6,no.1(January 24, 2012)
- ISSN：1936-086X
文摘
Using a single-molecule fluorescence approach, the time series of catalytic events of an enzymatic reaction can be monitored, yielding a sequence of fluorescent 鈥渙n鈥? and 鈥渙ff鈥?states. An accurate on/off-assignment is complicated by the intrinsic and extrinsic noise in every single-molecule fluorescence experiment. Using simulated data, the performance of the most widely employed binning and thresholding approach was systematically compared to change point analysis. It is shown that the underlying on- and off-histograms as well as the off-autocorrelation are not necessarily extracted from the 鈥渟ignal鈥欌€?buried in noise. The shapes of the on- and off-histograms are affected by artifacts introduced by the analysis procedure and depend on the signal-to-noise ratio and the overall fluorescence intensity. For experimental data where the background intensity is not constant over time we consider change point analysis to be more accurate. When using change point analysis for data of the enzyme 伪-chymotrypsin, no characteristics of dynamic disorder was found. In light of these results, dynamic disorder might not be a general sign of enzymatic reactions.