Department of Chemical Sciences, Indian Institute of Science Education and Research Bhopal, ITI Campus (Gas Rahat) Building, Govindpura, Bhopal 462 023, Madhya Pradesh, India.
Phys Chem Chem Phys. 2011 Dec 7;13(45):20418-26. doi: 10.1039/c1cp21759c. Epub 2011 Oct 13.
The mechanism by which the protein Bovine Serum Albumin (BSA) undergoes unfolding induced by Guanidine Hydrochloride (GdHCl) and then the subsequent refolding brought in by many-fold dilution was studied by steady-state fluorescence, anisotropy, time resolved measurements and Circular Dichroism (CD) spectroscopy. CD data reveal that the protein attains a degree of extra rigidity at low concentrations of the denaturant, GdHCl, and this observation was correlated with other techniques used in this present work. The unfolding and refolding of BSA appear to proceed through intermediates and both the processes are sequential in nature. The intrinsic fluorescence from the tryptophan amino acid residue of BSA and another external fluorophore Nile Red was made use of in order to investigate the mechanisms of unfolding and refolding and we have conclusively proved that both these processes follow a reversible mechanism.
本研究采用稳态荧光、各向异性、时间分辨测量和圆二色性（CD）光谱法，研究了牛血清白蛋白（BSA）在盐酸胍（GdHCl）诱导下展开，然后通过多次稀释复性的机制。CD 数据表明，蛋白质在低浓度变性剂 GdHCl 下达到一定程度的额外刚性，这一观察结果与本工作中使用的其他技术相关联。BSA 的展开和复性似乎通过中间体进行，这两个过程在本质上是顺序的。BSA 中的色氨酸氨基酸残基的固有荧光和另一个外部荧光尼罗红被用来研究展开和复性的机制，我们已经明确证明这两个过程都遵循可逆机制。
