本文關(guān)鍵詞：人唾液蛋白質(zhì)組的納升液相色譜-高分辨串聯(lián)質(zhì)譜鑒定及高豐度蛋白質(zhì)分析　出處：《質(zhì)譜學(xué)報(bào)》2016年05期 　論文類型：期刊論文

  更多相關(guān)文章： 唾液 蛋白質(zhì)組 納升液相色譜 高分辨質(zhì)譜 信號(hào)強(qiáng)度 高豐度蛋白質(zhì)

【摘要】：常規(guī)的定性分析質(zhì)譜數(shù)據(jù)能否以及如何應(yīng)用于蛋白質(zhì)組學(xué)的相對(duì)定量分析是實(shí)際工作中經(jīng)常遇到的問(wèn)題。本研究采用一維納升液相色譜-高分辨串聯(lián)質(zhì)譜(nano-HPLC-Triple TOF 5600)法鑒定人唾液蛋白質(zhì)組,并分析其基本組成,以探討質(zhì)譜數(shù)據(jù)與唾液中高豐度蛋白質(zhì)的相關(guān)性。本實(shí)驗(yàn)共鑒定了6 044條特異性酶切肽段,歸屬于α-淀粉酶1、碳酸酐酶6、血清白蛋白、粘蛋白、半胱氨酸蛋白酶抑制劑和免疫球蛋白等各種已知的高豐度蛋白質(zhì)在內(nèi)的521個(gè)蛋白,這為一維納升液相色譜分離條件下的唾液蛋白質(zhì)組的基本組成分析提供了參考。結(jié)果表明,僅用蛋白質(zhì)排序指標(biāo)Unused值表征蛋白質(zhì)的相對(duì)含量具有局限性,而蛋白質(zhì)排序、檢出肽段數(shù)目和肽段平均強(qiáng)度等綜合指標(biāo)與蛋白質(zhì)相對(duì)含量具有正相關(guān)性,即排名較后的蛋白質(zhì)均只有1條肽段被檢出,且肽段強(qiáng)度較低。這說(shuō)明,當(dāng)?shù)鞍踪|(zhì)相對(duì)含量差別較大時(shí),肽段的化學(xué)性質(zhì)和電離效率差異對(duì)質(zhì)譜信號(hào)的影響已不占主要地位,即可以認(rèn)為當(dāng)肽段檢出數(shù)目、鑒定覆蓋率和肽段強(qiáng)度均較低時(shí),蛋白質(zhì)的相對(duì)含量也較低。該結(jié)果可為利用Triple TOF 5600質(zhì)譜儀的定性鑒定數(shù)據(jù)進(jìn)行初步半定量判斷提供參考,也可為唾液蛋白質(zhì)組生物標(biāo)志物研究提供借鑒。
[Abstract]:Whether and how conventional qualitative analysis of mass spectrum data can be applied to the relative quantitative analysis of proteomics is a common problem in practice. Human saliva proteome was identified by nano-HPLC-Triple TOF 5600 method. In order to study the correlation between the mass spectrum data and the high abundance protein in saliva, 6 044 specific restriction peptides were identified, which belong to 偽 -amylase 1 and carbonic anhydrase 6. Serum albumin, mucin, cysteine protease inhibitors and immunoglobulin are among the 521 known high abundance proteins. This provides a reference for the basic composition analysis of saliva proteome under the condition of separation by liquid chromatography. The results show that this method can be used for the analysis of saliva proteome. It is limited to use protein sequencing index (Unused) to characterize the relative content of protein, while protein sequencing is limited. The number of peptides detected and the average strength of peptides were positively correlated with the relative content of protein, that is to say, only one peptide segment was detected in the bottom of the list, and the intensity of peptide segment was low. When the relative content of protein is different, the influence of the chemical properties and ionization efficiency of peptides on the mass spectrum signal is no longer dominant, that is to say, the number of peptides detected can be considered. When the identification coverage and peptide intensity were low. The relative content of protein is also relatively low. The results can provide a reference for the preliminary semi-quantitative determination by using the qualitative identification data of Triple TOF 5600 mass spectrometer. It can also provide reference for the study of salivary proteome biomarkers.
【作者單位】： 吉林大學(xué)中日聯(lián)誼醫(yī)院神經(jīng)內(nèi)科;吉林大學(xué)白求恩醫(yī)學(xué)部臨床醫(yī)學(xué)院;吉林大學(xué)中日聯(lián)誼醫(yī)院血管外科;深圳大學(xué)生命科學(xué)學(xué)院深圳市海洋生物資源與生態(tài)環(huán)境重點(diǎn)實(shí)驗(yàn)室;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(31470804) 吉林省科技廳項(xiàng)目(20130727029YY,20150204057SF) 深圳市科技項(xiàng)目(JSGG20140703163838793)資助
【分類號(hào)】：R446.1;O657.63
【正文快照】： Proteomic Identification and High Abundant Proteins Analysisof Human Saliva by Nano-Liquid Chromatography-HighResolution Tandem Mass SpectrometrySHI Kai 1,KONG Xiang-yi 2,DU Jian-shi 3,XU Jin-ling4,LI Shui-ming4,WANG Yong4,ZHAO Qing1唾液中的蛋白質(zhì)和多肽

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