Real-Time NMR Monitoring of Protein-Folding Kinetics by a Recycle Flow System for Temperature Jump

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• 作者：Kazuhiko Yamasaki ; Yuji Obara ; Manabu Hasegawa ; Hideki Tanaka ; Tomoko Yamasaki ; Tsuyoshi Wakuda ; Michiya Okada ; Takamitsu Kohzuma
• 刊名：Analytical Chemistry
• 出版年：2013
• 出版时间：October 15, 2013
• 年：2013
• 卷：85
• 期：20
• 页码：9439-9443
• 全文大小：284K
• 年卷期：v.85,no.20(October 15, 2013)
• ISSN：1520-6882

文摘

An NMR method was developed that allows for real-time monitoring of reactions (on the order of seconds) induced by a temperature jump. In a recycle flow system, heating and cooling baths were integrated, with the latter inside the NMR probe. A refolding reaction of ribonuclease A was triggered by rapid cooling and monitored by a series of NMR measurements over 12 s. Data were processed by principal component analysis, in which a factor related to the structural change with an exponential rate constant of 0.2鈥?.7 s^鈥? was successfully separated from factors related to baseline instability and/or noise. Temperature dependency of the rate constant revealed the entropy-driven formation of the transition state of the refolding reaction.