摘要
目的:分析Jurkat T细胞质膜蛋白质组成,并对这些质膜蛋白的生理学过程和功能进行初步分析,为进一步研究Jurkat T细胞膜蛋白功能奠定基础。方法:首先采用差异密度梯度离心法提取Jurkat T细胞膜蛋白,然后将提取出的膜蛋白根据分子量大小通过SDS-PAGE进行初步分离,再进一步将分离出的蛋白条带切下进行胶内酶解,酶解后的肽段通过液相-芯片-离子阱质谱技术进行鉴定和生物信息学分析,建立Jurkat T细胞质膜蛋白全谱图,并进一步通过GO(Gene Ontology)对这些质膜蛋白进行功能分析。结果:成功提取了Jurkat T细胞的膜总蛋白,并建立了Jurkat T细胞质膜蛋白全谱图,共鉴定出618个质膜蛋白,经GO注释分析,其中与结合功能相关的质膜蛋白有493个,与信号转导活性相关的有186个,具有酶催化活性的有166个,具有转运活性的有137个,有些还具有酶调节活性、结构分子活性或者运动活性等,功能尚不清楚的有49个。结论:通过差异密度梯度离心,结合一维SDS-PAGE和HPLC-CHIP-MS/MS,成功建立了Jurkat T细胞质膜蛋白全谱图,并通过GO注释,初步分析了这些蛋白的功能和生理学过程,为进一步研究Jurkat T细胞质膜蛋白的功能奠定了基础。
Objective: To analyze the components of Jurkat T cells plasma membrane protein by using one dimensional gel electrophoresis combined with HPLC-CHIP-MS/MS, and to study the physiological processand function of these proteins by GO (Gene Ontology) annotation for further study of Jurkat T cells membrane proteins. Methods: Differential density gradient centrifugation was used to extract membrane proteins from Jurkat T cells. After SDS-PAGE, all protein bands were cut off and gone through in-gel digestion. Then the obtained peptide fragments were identified by HPLC-CHIP-MS/MS.Bioinformatics was used to analyze the identified proteins. All plasma membrane proteins were picked out and analyzed by GO analysis. Results: Total membrane proteins of Jurkat T cells were successfully extracted, and the plasma membrane protein profile was established. After GO annotation, within the 618 membrane proteins identified, 493 were associated with binding function, 186 with signal transducer activity, 166 with catalytic activity, 137 with transporter activity, and others with enzyme regulator activity, motor activity, transcription regulator activity or antioxidant activity. Conclusions: The plasma membrane protein profile of Jurkat T cells was successfully established by differential density gradient centrifugation combined with one dimensional SDS-PAGE plus HPLC-CHIP-MS/MS and the physiological process and function of these proteins were preliminarily analyzed by GO annotation.
出处
《现代生物医学进展》
CAS
2009年第3期418-421,共4页
Progress in Modern Biomedicine
基金
国家自然科学基金(30500477)
国家高技术研究发展计划(863计划)(No.2006AA02A121)。