Ultraviolet Resonance Raman Spectroscopic Study on Regulatory Protein-Nucleic Acid Interactions

调节蛋白-核酸相互作用的紫外共振拉曼光谱研究

• 批准号: 62430004
• 负责人: HARADA Issei
• 金额: $ 15.81万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (A)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62430004/
• 关键词: Ultraviolet Resonance Raman Rpectroscopy; Regulatory Protein; Protein Nucleic Acid Interactions; Spectrum-Structure Relation; 蛋白ー核酸相互作用; DNA

The purpose of the research is threefold : construction of ultraviolet Raman apparatus, tabulation of UV Raman marker bands useful in structural study of proteins and nucleic acids, and UV resonance Raman (UVRR) spectroscopic study on cyclic AMP receptor protein (CRP) and the CRP-cAMP complex in solution.(1) UV Raman apparatusUV radiation is generated by a system consisting of a Nd : YAG laser (30 Hz), a harmonic generator (KDP or BBO), and an H_2-Raman shifter. Scattered light from a sample is dispersed (2nd order) by an f/6.8 80-cm double monochromator equipped with 600 g/mm gratings blazed at 500 nm and detected with a doubly intensified diode array of 700 pixels.(2) Table of UVRR marker bands and its applicationsUVRR spectra of peptides containing Pro, Trp, Tyr, Phe, and His, nucleosides, nucleotides, and the related compounds have been studied extensively and a list of useful structural marker bands is tabulated. Good-quality UVRR spectra of Cu, Zu-superoxide dismutase and bacteriorhodopsin have been recorded for the first time, and some pieces of structural information on the proteins are obtained on the basis of the table.(3) Structure of CRP and CRP-cAMP in dilute solutionUpon uptake of a cAMP molecule in one of the two subunits of CRP, drastic change occurs in the beta-sheet structure of the other subunit as well as the bound subunit. The adenine-ribose bond conformation is syn in the bound cAMP and the binding form is stabilized by H-bonding at N7 and C6NH_2 with side chains of the protein.

本研究的目的包括三个方面：紫外拉曼装置的建立，用于蛋白质和核酸结构研究的紫外拉曼标记谱带的列表，以及对溶液中环AMP受体蛋白(CRP)和CRP-cAMP络合物的紫外共振拉曼(UVRR)光谱研究。(1)紫外拉曼装置紫外拉曼辐射由Nd：YAG激光器(30 Hz)、谐波发生器(KDP或BBO)和H_2-拉曼移位器组成的系统产生。样品的散射光由f/6.8 80 cm双单色仪分散(二级)，该双单色仪配备600 g/mm的光栅，在500 nm处闪耀，并用700像素的双增强二极管阵列进行检测。(2)UVRR标记谱带及其应用首次记录了铜、锌超氧化物歧化酶和细菌视紫红质的高质量UVRR谱，并在此基础上获得了蛋白质的一些结构信息。(3)CRP和CRP-cAMP在稀溶液中的结构当CRP的两个亚基中的一个摄取cAMP分子时，另一个亚基和结合亚基的β-折叠结构发生了剧烈的变化。结合的cAMP中腺嘌呤-核糖键构象为SYN，结合形式由蛋白质侧链上的N7和C6NH2上的氢键稳定。

项目成果

T. Miura, H. Takeuchi, and I. Harada: "Tryptophan Raman Bands Sensitive to Hydrogen Bonding and Side Chain Conformation" J. Raman Spectrosc.20. 667-671 (1989)

T. Miura、H. Takeuchi 和 I. Harada：“色氨酸拉曼谱带对氢键和侧链构象敏感”J. Raman Spectrosc.20。

S. Hashimoto, S. Ohsaka, H. Takeuchi, and I. Harada: "Ultraviolet Resonance Raman Spectra of Cu, Zu-Superoxide Dismutase : Detection of an Imidazolate Bridge between the Metal Ions in Solution" J. Am. Chem. Soc.111. 8926-8928 (1989)

S. Hashimoto、S. Ohsaka、H. Takeuchi 和 I. Harada：“Cu、Zu 超氧化物歧化酶的紫外共振拉曼光谱：溶液中金属离子之间咪唑酯桥的检测”J. Am。

Takashi Miura: "Tryptophan Raman Bands Sensitive to Hydrogen Bonding and SideーChain Conformation" Journal of Raman Spectroscopy. 20. 667-671 (1989)

Takashi Miura：“色氨酸拉曼带对氢键和侧链构象敏感”拉曼光谱学杂志 20. 667-671 (1989)

Hideo Takeuchi: "Raman Bands of NーDeuterated Histidinium as Markers of Conformation and Hydrogen Bonding" Journal of Raman Spectroscopy. 22. (1991)

Hideo Takeuchi：“N-氘化组氨酸的拉曼带作为构象和氢键的标记”拉曼光谱学杂志 22。（1991）

Takashi Miura: "Tryptophan Raman Bands Sensitive to Hydrogen Bonding and SideーChain Conformation" J.Raman Spectrosc.20. 667-671 (1989)

Takashi Miura：“色氨酸拉曼带对氢键和侧链构象敏感”J.Raman Spectrosc.20 (1989)。