RELATION BETWEEN THE HIGHER ORDER STRUCTURAL CHANGES OF PROTEIN AND FUNCTION ABNORMARITY OF ABNORMAL HEMOGLOBIN

蛋白质高阶结构变化与异常血红蛋白功能异常的关系

• 批准号: 07670140
• 负责人: NAGAI Masako
• 金额: $ 1.54万
• 依托单位: KANAZAWA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670140/
• 关键词: Hemoglobin; Abnormal hemoglobin; Functinal abnormarity; UV resonance Raman; Circular dichroism; Higher order structure; Phosphotyrosine; 蛋白の高次構造変化; 紫外共鳴ラマン散乱; 円二色性

It has been recently reported that resonance Raman spectra excited in the UV region at 200-240 can explore the environmental and hydrogen bonding changes of tryptophan (Trp) and tyrosine (Tyr) residues of proteins. we have demonstrated here the quaternary-structural dependent features for Tyr and Trp residues in alpha_1beta_2 interface of hemoglobin (Hb) from 235-nm excited UV resonance Raman (UVRR) spectra. Deoxy Hb A (T-form) showd a UVRR spectrum distinctly different from those of the ligated Hbs (R-form) including osyHb, COHb and NOHb.To characterize the spectral changes of Trp-beta37 at alpha_1beta_2 interface due to the quaternary structure transition, the UVRR spectra of Hb A were compared with the corresponding spectra of Hb Hirose (Trp-beta37->Ser). Comparison of the Hb A - Hb Hirose difference spectra in oxy and deoxy states revealed that the oxygenation- induced changes of Trp RR bands arose mostly from Trp-beta37.The UVRR spectral contribution of alpha42Tyr, Which is located in the "swhitch" region of the alpha_1beta_2 interface and forms an H-bond with the carboxylate side chain of beta99 Asp only in the T-state, was deduced for each of the deoxy- and CO-forms by subtracting the spectra of Hb alpha Y42H from those of Hb A.This suggested that alpha42Tyr is responsible for the frequency shift of Y8a (1617cm^<-1>) and Y9a (1179cm^<-1>) of the Tyr RR bands of Hb alphaY42H upon quaternary structure change are alike.The extent of the oxidation of Hb M Sakatoon and Hb M Boston in the patients blood was determined by measurement of the intensity of EPR signal at g=6.0 for the normal subunits, g=6.7for the mutant subunit of Hb M Saskatoon, and g=6.3 for those of Hb M Boston. About 50% and 76% of mutant subunits in Hb M Boston and Hb M Sakatoon remained reduced in the fresh blood, respectively.

最近报道，在200-240的紫外区激发的共振拉曼光谱可以探索蛋白质色氨酸(Trp)和酪氨酸(Tyr)残基的环境和氢键变化。我们从235 nm激发的紫外共振拉曼(UVRR)光谱中证明了血红蛋白(Hb)α_1β_2界面Tyr和Trp残基的四元结构依赖特征。脱氧Hb A(T型)的UVRR谱与连接的HbS(R型)包括osyHb、COHb和NOHb的UVRR谱明显不同。为了表征由于四元结构转变而导致的α_1beta_2界面色氨酸-β37的光谱变化，比较了Hb A的UVRR谱和Hb Hiose(Trp-beta37-&GT；Ser)的相应光谱。比较Hb A-Hb在氧态和脱氧态的差谱表明，氧化引起的Trp RR谱带的变化主要来自Trp-Beta37。通过从Hb A的谱中减去HbαY42H的光谱，推测α42Tyr对UVRR谱的贡献是通过减去HbαY42H的光谱而得到的，α42Tyr位于α_1β_2界面的“swhitch”区，仅在T态与Beta99 Asp的羧基侧链形成H键。)和Y9a(1179 cm~(-1))和Y9a(1179 cm~(-1)~(-1))。通过测定正常亚基g=6.0、突变亚基g=6.7和Hb M Boston亚基g=6.3时Hb M Sakatoon和Hb M Boston的氧化程度，确定患者血液中Hb M Sakatoon和Hb M Boston的氧化程度。Hb M Boston和Hb M Sakatoon分别有约50%和76%的突变亚基在新鲜血液中保持还原。

项目成果

Nagai,M.: "Ultraviolet resonance Raman studies of hemoglobin quaternary structure using a tyrosine-42 mutant:changes in the α_1β_2 subunit interface upon the T→R transition." Journal of Molecular Structure. 379. 65-75 (1996)

Nagai, M.：“使用酪氨酸 42 突变体对血红蛋白四级结构进行紫外共振拉曼研究：T→R 转变时 α_1β_2 亚基界面的变化。”《分子结构杂志》379. 65-75 (1996)。

Nagai, M.: "Ultraviolet resonance Raman studies of hemoglobin quaternary structure using a tyrosine alpha42 mutant." J.Mol.Struct.379. 65-75 (1996)

Nagai, M.：“使用酪氨酸 alpha42 突变体对血红蛋白四级结构进行紫外共振拉曼研究。”

Nagai,M.: "Ultraviolet resonance Raman studies of quaternary structure of hemoglobin using a tryptophan β37 mutant." The Journal of Biological Chemistry. 270. 1636-1642 (1995)

Nagai, M.：“使用色氨酸 β37 突变体进行血红蛋白四级结构的紫外共振拉曼研究。”《生物化学杂志》270。1636-1642 (1995)

Nagai, M.: "Ultraviolet resonance Raman studies of quaternary structure of hemoglobin using a tryptophan beta37 mutant." J.Biol.Chem.270. 1636-1642 (1995)

Nagai, M.：“使用色氨酸 beta37 突变体对血红蛋白四级结构进行紫外共振拉曼研究。”

Nagai,M.: "Ultraviolet resonance Raman studies of quaternary structure of hemoglobin using a tryptophan β37 mutant." J.Biol.Chem.270(4). 1636-1642 (1995)

Nagai, M.：“使用色氨酸 β37 突变体对血红蛋白四级结构进行紫外共振拉曼研究。”J.Biol.Chem.270(4) (1995)。