Demchenko A P
Department of Biophysics, A.V. Palladin Institute of Biochemistry, Kiev, Ukraine.
Biochim Biophys Acta. 1994 Dec 14;1209(2):149-64. doi: 10.1016/0167-4838(94)90179-1.
With the advent and development of time-resolved spectroscopic techniques and substantial progress in understanding of photophysical and photochemical phenomena, a new goal may be achieved: modeling of biochemical reaction or its elementary step by a photochemical event occurring within the probe, bound to a protein molecule. The probe may be located in a well-determined site of the protein matrix and report on the modulation of the reaction rate by the matrix and by the surrounding solvent, or by interactions in multiprotein complexes and in biomembranes. The advantages of this approach are obvious: in contrast to ordinary biochemical reaction, the excited-state reaction may be started by a short light pulse, and its kinetics may be observed directly with high resolution in time. In addition, if the reaction rate is influenced by the dynamics of the protein matrix, these dynamics may be studied simultaneously with the reaction, by using the same or a similar probe and within the same time range. In this review, the prospects for application of probes exhibiting electron transfer, proton transfer, molecular rotations and isomerizations are presented and discussed. The general problem of photochemical modeling of biochemical reactions is discussed. This modeling may result in deeper understanding of enzyme catalyzed reaction mechanisms.
随着时间分辨光谱技术的出现与发展以及在光物理和光化学现象理解方面取得的重大进展，一个新的目标或许能够实现：通过与蛋白质分子结合的探针内部发生的光化学事件对生化反应或其基本步骤进行建模。该探针可定位在蛋白质基质的特定位置，并报告基质、周围溶剂、多蛋白复合物及生物膜中的相互作用对反应速率的调节情况。这种方法的优势显而易见：与普通生化反应不同，激发态反应可由短光脉冲引发，其动力学可在时间上以高分辨率直接观测。此外，如果反应速率受蛋白质基质动力学影响，那么可通过使用相同或类似探针并在相同时间范围内，与反应同时研究这些动力学。在本综述中，将介绍并讨论呈现电子转移、质子转移、分子旋转和异构化的探针的应用前景。还将讨论生化反应光化学建模的一般问题。这种建模可能会加深对酶催化反应机制的理解。
