【摘要】：離子淌度質(zhì)譜(ion mobility mass spectrum,IM-MS)即將離子淌度分離技術(shù)與質(zhì)譜聯(lián)用的一項(xiàng)新型分析方法,已成為一種強(qiáng)有力的結(jié)構(gòu)生物學(xué)分析工具,能同時(shí)測(cè)定分析物的結(jié)構(gòu)、拓?fù)鋵W(xué)結(jié)構(gòu)、動(dòng)態(tài)過程以及組成等。本論文以膜聯(lián)蛋白A5及其同質(zhì)二聚體diannexin和維生素B12結(jié)合蛋白F為材料,探究不同配體如鈣離子和維生素B12對(duì)蛋白質(zhì)結(jié)構(gòu)的影響。由電噴霧電離產(chǎn)生的蛋白質(zhì)離子在離子淌度質(zhì)譜儀的漂移管中與氣體發(fā)生碰撞而導(dǎo)致蛋白質(zhì)被活化,促使其構(gòu)象失去天然狀態(tài)發(fā)生去折疊等變化,且離子在漂移管中會(huì)根據(jù)他們的質(zhì)量、電荷、大小被逐漸分離。在本論文中我們采用Waters Synapt G2-Si HDMS質(zhì)譜儀采集離子的漂移時(shí)間,并經(jīng)過許多已知理論碰撞橫截面積的蛋白質(zhì)標(biāo)準(zhǔn)品做校正后得到離子天然構(gòu)象與去折疊狀態(tài)的碰撞橫截面積。實(shí)驗(yàn)發(fā)現(xiàn):未受碰撞活化的離子,其測(cè)得的碰撞橫截面積與預(yù)期的處于天然狀態(tài)的蛋白質(zhì)的碰撞橫截面積非常一致。而當(dāng)碰撞能量增加,離子被活化導(dǎo)致構(gòu)象發(fā)生變化,且構(gòu)象去折疊的程度與蛋白質(zhì)復(fù)合物分子內(nèi)或分子間的非共價(jià)相互作用的穩(wěn)定性有關(guān)。以上結(jié)果表明蛋白質(zhì)與配體結(jié)合后能使其在氣相中的結(jié)構(gòu)穩(wěn)定性大大增強(qiáng)。
[Abstract]:Ion mobility mass spectrometry (ion mobility mass spectrum,IM-MS), which combines ion mobility separation with mass spectrometry, has become a powerful tool for structural biological analysis, which can simultaneously determine the structure of analytes. Topological structure, dynamic process and composition, etc. In this paper, the effects of different ligands, such as calcium ion and vitamin B12, on protein structure were investigated by using membrane protein A5 and its homodimer diannexin and vitamin B12 binding protein F as materials. The protein ion produced by electrospray ionization collides with the gas in the drift tube of the ion mobility mass spectrometer, which causes the protein to be activated, causing its conformation to lose its natural state and unfold and so on. And ions in the drift tube will be gradually separated according to their mass, charge, and size. In this paper, we use Waters Synapt G2-Si HDMS mass spectrometer to collect the drift time of ion, and after correction of many protein standards with known theoretical collision cross-sectional area, we obtain the collision cross-sectional area of ion natural conformation and unfolding state. It is found that the collision cross-sectional area measured by the ion which is not activated by collision is in good agreement with the expected collision cross-sectional area of the protein in the natural state. When the collision energy increases, the ion activation causes the conformation change, and the degree of conformation unfolding is related to the stability of intramolecular or intermolecular noncovalent interactions of protein complexes. The above results indicate that the structure stability of the protein in gas phase can be greatly enhanced by binding the protein with ligand.
【學(xué)位授予單位】：南京理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：O657.63;TQ931

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 ;Identification of Bovine Casein Phosphorylation Using Titanium Dioxide Enrichment in Combination with Nano Electrospray Ionization Tandem Mass Spectrometry[J];Journal of Integrative Agriculture;2012年03期

2 張立勇,趙曉航,吳e，

本文編號(hào)：2453496