Probing protein electrostatics with a synthetic fluorescent amino acid
Probing protein electrostatics with a synthetic fluorescent amino acid
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DOI：
10.1126/science.1069346
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发表时间：
2002-05-31
期刊：
影响因子：
56.9
通讯作者：
Jan, LY
中科院分区：
文献类型：
--
作者：
Cohen, BE;McAnaney, TB;Jan, LY
静电几乎影响蛋白质结构和活性的所有方面，对于主要功能是稳定电荷的蛋白质尤其重要。在这里，我们介绍了一种荧光氨基酸 Aladan，它可以探测蛋白质多个位点的静电特性。 Aladan对其周围环境的极性异常敏感，并且可以在可溶性蛋白和膜蛋白中选择性地掺入埋藏和暴露的位点。对 G 蛋白 B1 结构域中不同埋藏和暴露位点的 Aladan 残基的稳态和时间分辨荧光测量表明，其内部是极性和异质的。
Electrostatics affect virtually all aspects of protein structure and activity and are particularly important in proteins whose primary function is to stabilize charge. Here we introduce a fluorescent amino acid, Aladan, which can probe the electrostatic character of a protein at multiple sites. Aladan is exceptionally sensitive to the polarity of its surroundings and can be incorporated site-selectively at buried and exposed sites, in both soluble and membrane proteins. Steady-state and time-resolved fluorescence measurements of Aladan residues at different buried and exposed sites in the B1 domain of protein G suggest that its interior is polar and heterogeneous.