Two dimensional near-infrared-infrared correlation spectroscopy studies of protein hydration and denaturation

蛋白质水化和变性的二维近红外-红外相关光谱研究

• 批准号: 11640516
• 负责人: OZAKI Yukihiro
• 金额: $ 2.18万
• 依托单位: Kwansei Gakuin University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11640516/
• 关键词: Two-Dimensional Infrared Spectroscopy; Infrared Spectroscopy; Near-Infrared Spectroscopy; Protein; Hydration; Denaturation

Attenuated total reflection (ATR)/infrared (IR) spectra were measured for human serum albumin (HSA) in aqueous solutions over a pH range of5.0-3.2.Generalized two-dimensional (2D) correlation analysis was applied to the amide I region of the spectra to investigate pH-dependent changes in secondary structures and in hydrogen bondings of side chains of HSA. The synchronous and asynchronous 2D spectra were generated from the pH-dependent spectral variations for the three states of HSA, the N isomeric form (pH 5.0-4.4), the N-F transition (pH 4.6-3.8), and the F isomeric form (pH 3.8-3.0). The most interesting finding in the 2D spectra is identification offour bands at 1740, 1715, 1705, and 1696 cm1 due to a OO stretching mode of free and hydrogen bonded (weak, medium, and strong) COOH groups ofHSA. "FT-NIR spectra of HSA at different concentration in aqueous solutions have been measured from 45-80 ーC. Concentration dependent 2D correlation analysis at different temperature were applied to … More these spectra in two range 5500-7500 cm1 and 4200-4900 cm"1, respectively. Absorption bands arising from the combination vibrations of O-H stretching in water dominate the NIR spectra of the aqueous solution, especially at the 5500-7500 cm'1 region. The hydration and inter-, intra-molecular interaction changes, mainly due to the variation of the hydrogen bonding between different molecular units have been provided by the 2D-correlation patterns in thisrange. "Molten globule-like states of ovalbumin (OVA) in acid aqueous solutions have been investigated by generalized 2D FT-NIR correlation spectroscopy. This new method allows us to explore in changes in hydration and secondary structure simultaneously.FT-NIR spectra were measured for OVA aqueous solutions with concentrations of 1, 2, 3, 4 and 5 wt% over a pH range of2.4-5.4.Concentration-perturbed 2D correlation spectra were calculated for'the spectra in the 4850-4200 cm1 and 7500-5350 cm"1 regions at different pH. The 2D-NIR synchronous spectrum in the 4850-4200 cm"1 region shows a significant change upon going from pH 5.4 to pH 3.6.An autopeak at 4265 cm"1 due to a combination of symmetric CH2 stretching mode and CH2 bending mode of side chains seen at pH 5.0 disappears completely in the synchronous spectrum at pH 3.6.This suggests that some amino acid residues of OVA are subjected to micro environmental changes with decreasing pH." Less

测定了pH值为5.0 ~ 3.2的水溶液中人血清白蛋白（HSA）的衰减全反射(ATR)/红外（IR）光谱。利用广义二维（2D）相关分析对光谱的酰胺I区进行分析，研究了HSA的二级结构和侧链氢键的ph依赖性变化。根据HSA三种形态(N-异构体形态（pH 5.0 ~ 4.4）、N-F跃迁形态（pH 4.6 ~ 3.8）和F异构体形态（pH 3.8 ~ 3.0）的pH依赖谱变化，生成同步和异步二维光谱。在二维光谱中，最有趣的发现是在1740、1715、1705和1696 cm2处发现了四个波段，这是由于hsa的自由和氢键（弱、中、强）COOH基团的OO拉伸模式。在45 ~ 80℃范围内测量了水溶液中不同浓度的HSA的FT-NIR光谱。分别在5500 ~ 7500 cm / 1和4200 ~ 4900 cm / 1两个范围内对这些光谱进行了不同温度下浓度相关分析。水溶液的近红外光谱主要是由O-H在水中拉伸的组合振动引起的吸收带，特别是在5500-7500 cm'1区域。水合作用和分子间、分子内相互作用的变化，主要是由于不同分子单元之间氢键的变化，在此范围内提供了二维相关模式。用广义二维FT-NIR相关光谱法研究了卵白蛋白（OVA）在酸性水溶液中的熔融球状态。这种新方法使我们能够同时探索水化和二级结构的变化。在2.4-5.4的pH范围内，测量了浓度为1,2,3,4,5 wt%的OVA水溶液的FT-NIR光谱。计算了不同pH值下4850 ~ 4200 cm / 1和7500 ~ 5350 cm / 1区域的二维浓度摄动相关光谱。从pH 5.4到pH 3.6, 4850 ~ 4200 cm / 1区域的二维近红外同步光谱发生了显著变化。在pH 5.0时，由于侧链的对称CH2拉伸模式和CH2弯曲模式的结合，在4265 cm“1处出现了自峰，但在pH 3.6时，这种自峰完全消失了。这表明OVA的一些氨基酸残基随着ph值的降低而受到微环境变化的影响

项目成果

Y.Wu, K.Murayama, and Y.Ozaki: "Two-Dimensional Infared Spectroscopy and Principle Component Analysis studies of the Secondary Structure and Kinetics of Hydrogen-Deuterrium Exchange of Human and sreum Alubmin"J. Phys. Chem. B. 105. 6251-6259 (2001)

Y.Wu、K.Murayama 和 Y.Ozaki：“人体和血清白蛋白氢-氘交换的二级结构和动力学的二维红外光谱和主成分分析研究”J。

B.Czarnik-Matusewicz, K.Murayama, Y.wu, and Y.Ozaki: "Two-Dimensional Attenuated Total Reflection/Infared Correlation Spectrospy of Adsorption-Induced and Concentration-Dependent Spectral Variations of β-Lactogllobulin in Aqueous Solutions"J. Phys. Chem.

B.Czarnik-Matusewicz、K.Murayama、Y.wu 和 Y.Ozaki：“水溶液中 β-乳球蛋白吸附诱导和浓度依赖性光谱变化的二维衰减全反射/红外相关光谱”J。物理化学。

K.Murayama, Y.Wu.B.Czarnik-Matusewicz, and Y, Ozaki: "Two-Dimensional/Attenuated Total Reflection Infared Correlation Apectroscopy Studies on Secondary Structural Changes in Human Serum Slubim in Aqueous Solutions : pH-Dependent Structural Changes in Soco

K.Murayama、Y.Wu.B.Czarnik-Matusewicz 和 Y, Ozaki：“二维/衰减全反射红外相关光谱研究水溶液中人血清 Slubim 的二次结构变化：Soco 中 pH 依赖性结构变化

Y.M.Jung,B.Czarnik-Matuseuicz Y.Ozaki: "Two-Dimensional Infrated, Two-Dimensional Raman and Two-Dimensional Infrared and Raman Heterospectral Correlation Studies of Secondary structure of β-Lactoglobulin in Buffer Solutions"J.Phys.Chem.. 140・32. 7812-7817

Y.M.Jung、B.Czarnik-Mateuicz Y.Ozaki：“缓冲溶液中 β-乳球蛋白二级结构的二维红外、二维拉曼和二维红外和拉曼异谱相关研究”J.Phys.Chem.. 140・32。7812-7817

B.Czarnik-Matusewicz, K.Murayama, Y.Wu, Y.ozaki: "Two-Dimensional Attenuated Total Reflection/Intrared Correlation Spectroscopy of Adsorption-Induced and Concentuation-Dependent Spectral Variations of β-Loctoglobulin in Aqueous"J.Phys.Chem. 104・32. 7803-7

B.Czarnik-Matusewicz、K.Murayama、Y.Wu、Y.ozaki：“二维衰减全反射/红外相关光谱分析水中 β-Loctoglobulin 的吸附诱导和浓度依赖性光谱变化”J.Phys。化学104・32。