School of Chemistry, University of East Anglia, Norwich Research Park, Norwich NR4 7TJ, United Kingdom.
Department of Physics, Stockholm University, SE-10691 Stockholm, Sweden.
J Phys Chem A. 2022 Feb 24;126(7):1158-1167. doi: 10.1021/acs.jpca.1c10628. Epub 2022 Feb 9.
The photophysics of green fluorescent protein (GFP) and red Kaede fluorescent protein (rKFP) are defined by the intrinsic properties of the light-absorbing chromophore and its interaction with the protein binding pocket. This work deploys photodissociation action spectroscopy to probe the absorption profiles for a series of synthetic GFP and rKFP chromophores as the bare anions and as complexes with the betaine zwitterion, which is assumed as a model for dipole microsolvation. Electronic structure calculations and energy decomposition analysis using Symmetry-Adapted Perturbation Theory are used to characterize gas-phase structures and complex cohesion forces. The calculations reveal a preponderance for coordination of betaine to the phenoxide deprotonation site predominantly through electrostatic forces. Calculations using the STEOM-DLPNO-CCSD method are able to reproduce absolute and relative vertical excitation energies for the bare anions and anion-betaine complexes. On the other hand, treatment of the betaine molecule with a point-charge model, in which the charges are computed from some common electron density population analysis schemes, show that just electrostatic and point-charge induction interactions are unable to account for the betaine-induced spectral shift. The present methodology could be applied to investigate cluster forces and optical properties in other gas-phase ion-zwitterion complexes.
绿色荧光蛋白(GFP)和红色 Kaede 荧光蛋白(rKFP)的光物理性质由吸光发色团的固有性质及其与蛋白质结合口袋的相互作用决定。本工作采用光解作用光谱法研究了一系列合成 GFP 和 rKFP 发色团的吸收谱,包括其作为游离阴离子以及与甜菜碱两性离子形成复合物时的吸收谱。甜菜碱两性离子被假定为偶极微溶的模型。采用对称性自适应微扰理论(Symmetry-Adapted Perturbation Theory)的电子结构计算和能量分解分析用于对气相结构和复合物内聚能进行特征化描述。计算结果表明,甜菜碱主要通过静电力优先配位到酚氧负离子去质子化位点。使用 STEOM-DLPNO-CCSD 方法进行的计算能够重现游离阴离子和阴离子-甜菜碱复合物的绝对和相对垂直激发能。另一方面,用点电荷模型处理甜菜碱分子,其中电荷是根据一些常见的电子密度布居分析方案计算得到的,表明仅仅静电和点电荷诱导相互作用无法解释甜菜碱引起的光谱位移。本方法可以应用于研究其他气相离子-两性离子复合物中的团簇力和光学性质。