Yadavalli Venkateswarlu, Nellaepalli Sreedhar, Subramanyam Rajagopal
Department of Biochemistry, School of Life Sciences, University of Hyderabad, Hyderabad, India.
Methods Mol Biol. 2011;684:159-70. doi: 10.1007/978-1-60761-925-3_14.
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.
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。