论文部分内容阅读
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 plasma membrane proteins of Jurkat T cells and to analyze the physiological processes and functions of these plasma membrane proteins, which will lay the foundation for further study on the membrane proteins of Jurkat T cells. Methods: Firstly, the membrane proteins of Jurkat T cells were extracted by differential density gradient centrifugation. Then, the extracted membrane proteins were separated by SDS-PAGE according to their molecular weight. The protein bands were further separated and gel- The digested peptides were identified and bioinformatically analyzed by liquid chromatography-on-chip-ion trap mass spectrometry. The full-length plasma membrane protein profiles of Jurkat T cells were established and further analyzed by GO (Gene Ontology) . Results: The total membrane protein of Jurkat T cells was successfully extracted and the whole plasma membrane protein profile of Jurkat T cells was established. A total of 618 plasma membrane proteins were identified and analyzed by GO. Among them, 493 There were 186 related to signal transduction activity, 166 with enzymatic activity, 137 with transport activity, and some also had enzyme regulatory activity, structural molecular activity, or locomotor activity, etc. The function was unclear There are 49. Conclusion: The full-scale plasma membrane protein profile of Jurkat T cells was successfully established by differential density gradient centrifugation combined with one-dimensional SDS-PAGE and HPLC-CHIP-MS / MS. The functional and physiological processes of these proteins were primarily analyzed by GO annotation , Which laid the foundation for further study of the function of Jurkat T cell plasma membrane protein.