人唾液蛋白质组的纳升液相色谱-高分辨串联质谱鉴定及高丰度蛋白质分析

发布时间：2018-01-13 09:05

本文关键词：人唾液蛋白质组的纳升液相色谱-高分辨串联质谱鉴定及高丰度蛋白质分析　出处：《质谱学报》2016年05期 　论文类型：期刊论文

【摘要】：常规的定性分析质谱数据能否以及如何应用于蛋白质组学的相对定量分析是实际工作中经常遇到的问题。本研究采用一维纳升液相色谱-高分辨串联质谱(nano-HPLC-Triple TOF 5600)法鉴定人唾液蛋白质组,并分析其基本组成,以探讨质谱数据与唾液中高丰度蛋白质的相关性。本实验共鉴定了6 044条特异性酶切肽段,归属于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剂和免疫球蛋白等各种已知的高丰度蛋白质在内的521个蛋白,这为一维纳升液相色谱分离条件下的唾液蛋白质组的基本组成分析提供了参考。结果表明,仅用蛋白质排序指标Unused值表征蛋白质的相对含量具有局限性,而蛋白质排序、检出肽段数目和肽段平均强度等综合指标与蛋白质相对含量具有正相关性,即排名较后的蛋白质均只有1条肽段被检出,且肽段强度较低。这说明,当蛋白质相对含量差别较大时,肽段的化学性质和电离效率差异对质谱信号的影响已不占主要地位,即可以认为当肽段检出数目、鉴定覆盖率和肽段强度均较低时,蛋白质的相对含量也较低。该结果可为利用Triple TOF 5600质谱仪的定性鉴定数据进行初步半定量判断提供参考,也可为唾液蛋白质组生物标志物研究提供借鉴。
[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 伪 -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.
【作者单位】：吉林大学中日联谊医院神经内科;吉林大学白求恩医学部临床医学院;吉林大学中日联谊医院血管外科;深圳大学生命科学学院深圳市海洋生物资源与生态环境重点实验室;
【基金】：国家自然科学基金项目(31470804) 吉林省科技厅项目(20130727029YY,20150204057SF) 深圳市科技项目(JSGG20140703163838793)资助
【分类号】：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唾液中的蛋白质和多肽

【相似文献】