基于电喷雾电离-捕集离子迁移谱-飞行时间质谱检测血红蛋白在不同pH值和醇溶液下的构象变化

引用本文: 张立昱, 唐科奇, 胡军. 基于电喷雾电离-捕集离子迁移谱-飞行时间质谱检测血红蛋白在不同pH值和醇溶液下的构象变化[J]. 分析化学, 2021, 49(8): 1335-1341. doi: 10.19756/j.issn.0253-3820.211021 shu

Citation: ZHANG Li-Yu, TANG Ke-Qi, HU Jun. Investigation of Conformational Changes of Hemoglobin at Different pH Values and in Different Alcohol Solutions by Electrospray Ionization-Trapped Ion Mobility Spectrometry-Time-of-Flight Mass Spectrometry[J]. Chinese Journal of Analytical Chemistry, 2021, 49(8): 1335-1341. doi: 10.19756/j.issn.0253-3820.211021 shu

基于电喷雾电离-捕集离子迁移谱-飞行时间质谱检测血红蛋白在不同pH值和醇溶液下的构象变化

宁波大学材料科学与化学工程学院, 质谱技术与应用研究院, 宁波 315211

通讯作者: 唐科奇,E-mail:tangkeqi@nbu.edu.cn; 胡军,E-mail:hujun@nbu.edu.cn

摘要: 蛋白质是重要的生物分子之一，具有很多重要的生物功能。大量的研究表明，蛋白质分子的功能与其分子结构和构象有关，因此，研究蛋白质分子的结构和变化是了解其生物功能和作用过程的主要途径之一。本研究利用质谱和离子迁移谱技术，研究了血红蛋白在不同酸碱度、不同浓度的醇溶液下构象变化情况。实验结果表明，血红蛋白在pH=4时会发生明显的结构改变，并观察到血红素脱落的现象。随着pH值降低，其离子迁移率进一步增大，说明随着溶液酸度逐渐增加，蛋白质解折叠的程度越来越大。通过观察离子迁移率的变化，推测了血红蛋白在不同pH值下的折叠情况，并研究了相同pH值条件下，携带不同电荷的血红蛋白离子的折叠情况，结果表明，所带电荷越多，其解折叠程度越大。此外，通过考察血红蛋白在不同浓度的醇溶液中血红素脱落的情况和离子迁移率，发现不同种类的醇对血红蛋白折叠状态的影响程度不同，如50%（V/V）甲醇溶液可导致血红蛋白折叠状态明显改变，但40%（V/V）乙醇溶液可导致几乎相同的结构变化。在高比例乙醇溶液中，血红蛋白分子会发生结构坍塌现象，使得血红蛋白在高比例醇溶液中的结构更加紧凑。

关键词:

English

Investigation of Conformational Changes of Hemoglobin at Different pH Values and in Different Alcohol Solutions by Electrospray Ionization-Trapped Ion Mobility Spectrometry-Time-of-Flight Mass Spectrometry

School of Materials Science and Chemical Engineering, Institute of Mass Spectrometry, Ningbo University, Ningbo 315211, China

Corresponding author: TANG Ke-Qi, ; HU Jun,

Received Date: 11 January 2021
Revised Date: 05 May 2021

Abstract: Protein, as one of the important biomolecules, plays important roles in many biological processes. Studies have shown that the function of the protein is closely related to its structure. Therefore, elucidating the structure changes of the protein in a given biological environment is one of the main ways to understand its biological function in a specific process. In this work, a high resolution mass spectrometry (MS) combined with ion mobility spectrometry (IMS) was used to study the conformation changes of hemoglobin under different conditions, such as different pH values and different solvents. The result showed that the structure of hemoglobin changed significantly at pH=4 with observed heme drop. Moreover, the ion mobility of the hemoglobin ions increased as the decrease of pH values, indicating the unfolding of the protein under the acidic solution conditions. By measuring the change of ion mobility, the unfolding of hemoglobin at different pH values was determined. Also the structural changes of hemoglobin ions as the change of the charge states at the same pH values were investigated and it was found that the structure of the hemoglobin became more extended as the increase of the protein charge state. In addition, it was found that the structure of the hemoglobin was also significantly affected by the type and the concentration of alcohol in the solvent. Specifically, the structure of hemoglobin in 50% methanol in water was about the same as the structure in 40% of ethanol aqueous solution. It was also found that the structure of hemoglobin collapsed in high concentration of alcohol solution, which made the structure of hemoglobin in high concentration of alcohol solution more compact.

Key words:

Hemoglobin
/ Electrospray ionization-trapped ion mobility spectrometry-time-of-flight mass spectrometry
/ Folding state
/ Conformation

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沈阳化工大学材料科学与工程学院沈阳 110142

基于电喷雾电离-捕集离子迁移谱-飞行时间质谱检测血红蛋白在不同pH值和醇溶液下的构象变化

通讯作者: 唐科奇,E-mail:tangkeqi@nbu.edu.cn; 胡军,E-mail:hujun@nbu.edu.cn

English

Investigation of Conformational Changes of Hemoglobin at Different pH Values and in Different Alcohol Solutions by Electrospray Ionization-Trapped Ion Mobility Spectrometry-Time-of-Flight Mass Spectrometry

Corresponding author: TANG Ke-Qi, ; HU Jun,

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基于电喷雾电离-捕集离子迁移谱-飞行时间质谱检测血红蛋白在不同pH值和醇溶液下的构象变化

通讯作者: 唐科奇,E-mail:tangkeqi@nbu.edu.cn; 胡军,E-mail:hujun@nbu.edu.cn

English

Investigation of Conformational Changes of Hemoglobin at Different pH Values and in Different Alcohol Solutions by Electrospray Ionization-Trapped Ion Mobility Spectrometry-Time-of-Flight Mass Spectrometry

Corresponding author: TANG Ke-Qi, ; HU Jun,

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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通讯作者: 陈斌, bchen63@163.com

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