Naseem Farah, Khan Rizwan Hasan
Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh-202002, India.
J Protein Chem. 2003 Nov;22(7-8):677-82. doi: 10.1023/b:jopc.0000008733.93463.ed.
Effect of increasing concentrations of two of the polyols, ethylene glycol (EG) and polyethylene glycol (PEG), was studied by near and far circular dichroism (CD), fluorescence emission spectroscopy, and binding of hydrophobic dye, 1-anilino-8-naphthalene sulfonic acid (ANS). Far-UV CD spectra show the transition of acid-unfolded trypsinogen from an unordered state to an intermediate state having ordered secondary structure. Interestingly, near-UV CD spectra show some amounts of stabilizing effect on the tertiary structure of the protein also. Tryptophan fluorescence studies indicate the change in the environment of the tryptophan residues on addition of EG and PEG. Maximum ANS binding occurs in presence of 80% EG and 90% PEG (v/v), suggesting the presence of an intermediate or molten globule-like state at high concentrations of the two polyols.
通过近圆二色性(CD)和远圆二色性、荧光发射光谱以及疏水性染料1-苯胺基-8-萘磺酸(ANS)的结合,研究了两种多元醇(乙二醇(EG)和聚乙二醇(PEG))浓度增加的影响。远紫外CD光谱显示酸解折叠的胰蛋白酶原从无序状态转变为具有有序二级结构的中间状态。有趣的是,近紫外CD光谱也显示出对蛋白质三级结构有一定程度的稳定作用。色氨酸荧光研究表明,添加EG和PEG后色氨酸残基的环境发生了变化。在80% EG和90% PEG(v/v)存在的情况下,ANS结合达到最大值,这表明在高浓度的这两种多元醇存在时,存在中间态或类熔球态。