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类囊体膜的蛋白质组学分析

Proteomic analysis of thylakoid membranes.

作者信息

Yadavalli Venkateswarlu, Nellaepalli Sreedhar, Subramanyam Rajagopal

机构信息

Department of Biochemistry, School of Life Sciences, University of Hyderabad, Hyderabad, India.

出版信息

Methods Mol Biol. 2011;684:159-70. doi: 10.1007/978-1-60761-925-3_14.

Abstract

Chlamydomonas is a model organism to study photosynthesis. Thylakoid membranes comprise several proteins belonging to photosystems I and II. In this chapter, we show the accurate proteomic measurements in thylakoid membranes. The chlorophyll-containing membrane protein complexes were precipitated using chloroform/methanol solution. These complexes were separated using two-dimensional gel electrophoresis, and the resolved spots were exercised from the gel matrix and digested with trypsin. These peptide fragments were separated by MALDI-TOF, and the isotopic masses were blasted to a MASCOT server to obtain the protein sequence. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF). The method discussed here would be a useful method for the separation and identification of thylakoid membrane proteins.

摘要

衣藻是研究光合作用的模式生物。类囊体膜包含几种属于光系统I和光系统II的蛋白质。在本章中,我们展示了类囊体膜中精确的蛋白质组学测量结果。使用氯仿/甲醇溶液沉淀含叶绿素的膜蛋白复合物。这些复合物通过二维凝胶电泳进行分离,从凝胶基质中切下分离出的斑点并用胰蛋白酶消化。这些肽片段通过基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF)进行分离,其同位素质量被输入到MASCOT服务器中以获得蛋白质序列。本文讨论的方法将是分离和鉴定类囊体膜蛋白的一种有用方法。

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