Department of Chemistry, University of Calcutta, Kolkata 700 009, India.
J Photochem Photobiol B. 2010 Dec 2;101(3):304-12. doi: 10.1016/j.jphotobiol.2010.07.016. Epub 2010 Aug 4.
The binding interactions of protein Bovine Serum Albumin (BSA) in its folding, unfolding and refolding states with proton transfer fluorescence probe 2-hydroxy-pyridine (2HP) and 5-(4-fluorophenyl)-2-hydroxypyridine (FP2HP) have been studied using steady state and time-resolved spectroscopy. The higher degree of spectral overlap between donor fluorescence and acceptor absorption is responsible for energy transfer from donor tryptophan to the acceptor probe and has shown remarkable sensitivity of these fluorophore for mapping the protein environment. During denaturation of BSA by guanidine hydrochloride, it shows two peaks of Trp-212 and Tyr-263. Reduction of fluorescence intensity of two peaks upon binding to the probes indicates that these probes not only bind with Trp-212 but also with Tyr-263. The steady state results are also confirmed by time-resolved studies.
采用稳态和时间分辨光谱法研究了蛋白质牛血清白蛋白（BSA）在折叠、展开和重折叠状态下与质子转移荧光探针 2-羟基吡啶（2HP）和 5-（4-氟苯基）-2-羟基吡啶（FP2HP）的结合相互作用。供体荧光与受体吸收之间更高的光谱重叠度负责从供体色氨酸到受体探针的能量转移，并表现出这些荧光团对蛋白质环境进行映射的显著灵敏度。在盐酸胍变性 BSA 时，它显示出色氨酸-212 和酪氨酸-263 的两个峰。结合探针后两个峰的荧光强度降低表明，这些探针不仅与色氨酸-212 结合，而且与酪氨酸-263 结合。稳态结果也通过时间分辨研究得到证实。
