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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >ULTRAFAST THERMALLY INDUCED UNFOLDING OF RNASE A
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ULTRAFAST THERMALLY INDUCED UNFOLDING OF RNASE A

机译：超热诱导RNASE A的折叠

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A temperature jump (T-jump) method capable of initiating thermally induced processes on the picosecond time scale in aqueous solutions is introduced. Protein solutions are heated by energy from a laser pulse that is absorbed by homogeneously dispersed molecules of the dye crystal violet. These act as transducers by releasing the energy as heat to cause a T-jump of up to 10 K with a time resolution of 70 ps, The method was applied to the unfolding of RNase A. At pH 5.7 and 59 degrees C, a T-jump of 3-6 K induced unfolding which was detected by picosecond transient infrared spectroscopy of the amide I region between 1600 and 1700 cm(-1). The difference spectral profile at 3.5 ns closely resembled that found for the equilibrium (native - unfolded) states. The signal at 16330 cm(-1), corresponding to the beta-sheet structure, achieved 15 +/- 2% of the decrease found at equilibrium, within 5.5 ns. However, no decrease in absorbance was detected until 1 ns after the T-jump. The disruption of beta-sheet therefore appears to be subject to a delay of approximate to 1 ns. Prior to 1 ns after the T-jump, water might be accessing the intact hydrophobic regions. [References: 58]
机译：介绍了一种能够在皮秒级的水溶液中引发热诱导过程的温度跳跃（T跃迁）方法。蛋白质溶液由来自激光脉冲的能量加热，该能量被染料结晶紫的均匀分散的分子吸收。它们通过释放能量作为热量而充当换能器，引起高达10 K的T跃迁，时间分辨率为70 ps。该方法应用于RNase A的展开。在pH 5.7和59摄氏度下，T跳跃的3-6 K诱导的解开，通过皮秒瞬态红外光谱对1600-1700 cm（-1）之间的酰胺I区进行检测。 3.5 ns处的光谱差异图非常类似于平衡状态（本征-展开）。在16330 cm（-1）处的信号（对应于β-折叠结构）在5.5 ns内达到了平衡时下降的15 +/- 2％。但是，直到T跃迁后1 ns才检测到吸光度降低。因此，β-sheet的破坏似乎要经历大约1 ns的延迟。在T跳之后的1 ns之前，水可能会进入完整的疏水区域。 [参考：58]

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