基于OFFGEL预分离二维液质联用技术在大鼠海马膜蛋白中的应用

以大鼠海马膜组分为研究对象,以等电聚焦OFFGEL为研究手段,结合反相液相色谱质谱联用技术,考察对酶切肽段的分离富集性能,同时结合DAVID分析工具对分离富集得到的酸性蛋白质进行的GO分析.结果表明,海马膜组分的酶切肽段经过OFFGEL分离后,得到24个不同馏分,每个馏分经过纳流液相色谱-质谱检测,发现有77% 的肽段专属于一个馏分,在酸性范围内的肽段等电点偏差较小,可以达到良好的聚焦效果;酸性蛋白的GO分析结果显示,质膜和物质的跨膜运输所占比例最大.OFFGEL作为一种预分离手段,可以应用于生物样品的大规模蛋白鉴定,在进一步蛋白质层面的生物功能研究中表现出良好的应用前景.

Study of Rat Hippocampus Membrane Proteins Using the Technology of Two-Dimensional High Performance Liquid Chromatography-Mass Spectrometry Based on OFFGEL

A study on the membrane components of rat hippocampus was presented. The isoelectric focusing OFFGEL combing with the reversed-phase liquid chromatography and mass spectrometry was adopted as the major tool to study the separation and enrichment capability of tryptic peptides. The database for annotation, visualization and integrated discovery(DAVID) was used to analyze the acidic proteins which were separated based on GO (gene ontology). The results indicate that there are 24 different fractions, each of which is identified by nano-reversed-phase liquid chromatography coupled with quadruple tandem time-of-flight mass spectropho-tometer (Q-TOF). About 77% of the identified peptides are found in only one fraction. Moreover, little deviations are observed at low pH value and it has a good focusing effect.