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功能化衍生試劑設(shè)計(jì)、合成及多肽標(biāo)記研究

發(fā)布時(shí)間:2018-07-29 19:04
【摘要】:發(fā)展新型質(zhì)譜兼容性的衍生試劑對于提高蛋白質(zhì)/多肽的質(zhì)譜離子化效率和深入的蛋白質(zhì)組研究具有重要的意義。基于此,本文設(shè)計(jì)、合成了五種新型的咪唑季銨類衍生試劑并考察了衍生化對質(zhì)譜鑒定效果的影響。 合成了五種新型的芳香季銨類衍生試劑,包括碘化1-(4-碘代丁烷)-3-丁基咪唑(IBBI)、溴化1-丙醛-3-丁基咪唑(PPBI)、溴化1-[3-[(2-碘代-1-乙氧基)]氨基丙基]-3-甲基咪唑(IPMI)、溴化1-[3-[(2-碘代-1-乙氧基)]氨基丙基]-3-丁基咪唑(IPBI)和溴化1-[3-[(2-碘代-1-乙氧基)]氨基丙基]-3-己基咪唑(IPHI),并進(jìn)一步采用質(zhì)譜和核磁共振進(jìn)行了衍生試劑結(jié)構(gòu)的確認(rèn)。 將衍生試劑IPBI應(yīng)用于半胱氨酸肽段的衍生和質(zhì)譜分析。肽衍生產(chǎn)物顯示了接近100%的標(biāo)記效率和高穩(wěn)定性。此外,肽段經(jīng)衍生后可同時(shí)提高肽段的質(zhì)譜離子化效率和質(zhì)譜帶電荷數(shù),,優(yōu)于常用的烷基化衍生試劑IAA的衍生效果和最近報(bào)道的衍生試劑IDA的衍生效果。 比較了試劑IPMI、IPBI和IPHI衍生對半胱氨酸肽段質(zhì)譜鑒定的影響。三種衍生試劑均顯示接近100%的衍生化標(biāo)記效率和衍生產(chǎn)物的高穩(wěn)定性、改進(jìn)的離子化效率和增加的質(zhì)譜帶電荷數(shù)。其中,IPHI具有最強(qiáng)的疏水性,可顯著提高肽段在MALDI-TOF MS質(zhì)譜中離子化。
[Abstract]:It is of great significance to develop a new type of derivative reagent compatible with MS for improving the ionization efficiency of protein / polypeptide mass spectrometry and for further proteomics research. Based on this, five new imidazole quaternary ammonium derivatives were designed and synthesized, and the effect of derivatization on the identification by mass spectrometry was investigated. Five new aromatic quaternary ammonium derivatives were synthesized. Including 1- (4-iodobutane) -3-Ding Ji imidazole (IBBI), brominated 1-propionaldehyde -3- Ding Ji imidazole (PPBI), bromination 1- [3- [(2-iodo-1-ethoxy)] -aminopropyl] -3-methylimidazole (IPMI), bromination 1- [3- [(2-iodo-1-ethoxy)] aminopropyl] -3-Ding Ji imidazole (IPBI) and 1- [(2-iodo-1-ethoxy)] aminopropyl] -3- Ding Ji imidazole (IPBI) The structure of the derivative reagent was further confirmed by (IPHI), with iodo-1-ethoxy] aminopropyl] -3-hexyl imidazole. The derivative reagent IPBI was applied to the derivatization and mass spectrometry of cysteine peptide. Peptide derivatives showed close to 100% labeling efficiency and high stability. In addition, the ionization efficiency of peptide segment and the charge number of mass spectrum can be increased simultaneously, which is superior to that of IAA, a common alkylation derivative, and the derivative effect of IDA, which has been reported recently. The effects of IPMI-IPBI and IPHI derivatives on the identification of cysteine peptide segment mass spectrometry were compared. All three kinds of derivatization reagents showed nearly 100% derivatization labeling efficiency, high stability of derivatives, improved ionization efficiency and increased mass spectrum charge number. The highest hydrophobicity was found in the MALDI-TOF MS mass spectrometry, which could significantly increase the ionization of the peptide segment in the MALDI-TOF MS mass spectrometry.
【學(xué)位授予單位】:河北大學(xué)
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2014
【分類號】:R914

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 喬曉強(qiáng);王蕊;張麗華;楊更亮;張玉奎;;肽段化學(xué)衍生技術(shù)及其在質(zhì)譜高靈敏度鑒定中的應(yīng)用進(jìn)展[J];分析化學(xué);2012年07期



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