Stability of the structure and redox state of ferricytochrome c during continuous changing of the hydration environment from dilute solution to condensed phase was studied based on real-time resonance Raman measurement. Our experimental results indicated that the structure and redox state of ferricytochrome c were sufficiently maintained during the entire desolvation process, which reflected that the stabilizing excluded-volume effect was stronger than the destabilizing crowder-protein interaction, at the case of globular mini-protein itself acting as the crowder. Rapid reduction of ferricytochrome c was observed at measurement sites at the time of solution to condensed phase transition. Similar phenomenon was observed on cytochrome c adsorbed on indium tin oxide surface. At present conditions, the rapid reduction was found to be derived mainly from thermal reduction under laser illumination and secondarily from photoreduction induced by the residual adsorbed ferrocyanide anions. After re-dissolved, the protein which had undergone sufficient desolvation recovered the structure and redox state in the initial solution phase, suggesting that the crowding effect in solution prevented large protein deformations observed in gas phase condition.

基于实时共振拉曼测量，研究了水合环境从稀溶液到浓缩相连续变化过程中亚铁色素c的结构和氧化还原状态的稳定性。我们的实验结果表明，在整个去溶剂化过程中，亚铁色素c的结构和氧化还原状态都得到了充分的维持，这表明在球状微蛋白自身的情况下，稳定的排阻体积作用比不稳定的拥挤蛋白相互作用更强。充当拥挤者。在溶液向缩合相转变时，在测量部位观察到亚铁细胞色素c的快速还原。在细胞色素c上观察到类似现象吸附在氧化铟锡表面上。在当前条件下，发现快速还原主要是由于激光照射下的热还原，其次是由于残留的吸附的亚铁氰化物阴离子引起的光还原。重新溶解后，经过充分去溶剂化处理的蛋白质在初始溶液阶段恢复了结构和氧化还原状态，这表明溶液中的拥挤效应阻止了在气相条件下观察到的大蛋白质变形。