Single-molecule fluorescence spectroscopy of enzyme conformational dynamics and cleavage mechanism
Single-molecule fluorescence spectroscopy of enzyme conformational dynamics and cleavage mechanism
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DOI：
10.1073/pnas.96.3.893
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发表时间：
1999-02-02
影响因子：
11.1
通讯作者：
Weiss, S
中科院分区：
文献类型：
--
作者：
Ha, TJ;Ting, AY;Weiss, S
荧光共振能量转移和荧光偏振各向异性用于研究葡萄球菌核酸酶的单分子。荧光标记的葡萄球菌核酸酶分子的分子内荧光共振能量转移和荧光偏振各向异性测量揭示了可能归因于毫秒时间尺度上的蛋白质构象动力学的波动的不同模式。分子间荧光共振能量转移测量提供了关于葡萄球菌核酸酶与单个底物分子的动态相互作用的信息。这里展示的实验方法应证明在单分子分辨率的蛋白质折叠和酶催化的研究中普遍有用。
Fluorescence resonance energy transfer and fluorescence polarization anisotropy are used to investigate single molecules of the enzyme staphylococcal nuclease. Intramolecular fluorescence resonance energy transfer and fluorescence polarization anisotropy measurements of fluorescently labeled staphylococcal nuclease molecules reveal distinct patterns of fluctuations that may be attributed to protein conformational dynamics on the millisecond time scale. Intermolecular fluorescence resonance energy transfer measurements provide information about the dynamic interactions of staphylococcal nuclease with single substrate molecules. The experimental methods demonstrated here should prove generally useful in studies of protein folding and enzyme catalysis at single-molecule resolution.