小分子探针对光敏黄蛋白结构和动力学的研究

Study on the Structure and Dynamics of Photoactive Yellow Protein Mutant M100A by a Small Probe Molecule

以二维红外光谱技术(2DIR)为主要的研究手段,以过渡金属羰基配合物CORM-3作为外源探针,研究了光敏黄蛋白(photoactive yellow protein,PYP)突变体M100A的超快结构动力学。结果表明,CORM-3以RuII(CO)2(RC)的形式稳定地结合在蛋白质的两个位点(H3和E81)上,并在红外光谱的羰基伸缩振动区域会产生四个吸收峰。2DIR结果表明两个结合位点存在不同的C≡O振动光谱扩散动力学,说明这两个结合位点处RC的水合程度以及蛋白质的局部结构环境不同。

We use two-dimensional infrared(2DIR)as a main experimental approach to study the ultrafast dynamics of the M100A mutant of photoactive yellow protein(PYP)with the aid of a transition metal carbonyl complex,CORM-3.Our results show that CORM-3binds to M100Ain the form of RuII(CO)2(RC)at two stable binding sites,H3and E81,generating four peaks in the C≡O stretching region of FTIR.2DIR show that two bindingsites have different C≡O vibrational spectral diffusion dynamics,indicating different hydration extent and different local protein structural environment.