Purification of fully active and crystallizable photosystem II from thermophilic cyanobacteria

Keisuke Kawakami, Jian Ren Shen

    Research output: Chapter in Book/Report/Conference proceedingChapter

    4 Citations (Scopus)

    Abstract

    Photosystem II (PSII) is a membrane protein complex which functions to catalyze light-induced water oxidation in oxygenic photosynthesis. Through the water-splitting reaction of PSII, light energy is converted into biologically useful chemical energy, and molecular oxygen is formed which transformed the atmosphere into an aerobic one and sustained aerobic life on the Earth. The PSII core complex from cyanobacteria consists of 17 transmembrane subunits and 3 extrinsic subunits with a total molecular mass of approximately 350 kDa per monomer, and PSII exists predominately in a dimeric form in vivo. This chapter describes the purification procedures leading to highly pure, homogenous, and highly active PSII core dimers from a thermophilic cyanobacterium, Thermosynechococcus vulcanus (T. vulcanus), that are used for successful crystallization and diffraction at atomic resolution. The purity and homogeneity of the PSII dimers thus obtained are characterized by absorption spectra, low-temperature fluorescence spectra, SDS-PAGE, clear native PAGE, blue native PAGE, gel filtration chromatography, and oxygen-evolving activity measurements. Finally, high-quality crystals obtained from the purified PSII dimers are shown.

    Original languageEnglish
    Title of host publicationMethods in Enzymology
    EditorsFraser Armstrong
    PublisherAcademic Press Inc.
    Pages1-16
    Number of pages16
    ISBN (Print)9780128163610
    DOIs
    Publication statusPublished - 2018

    Publication series

    NameMethods in Enzymology
    Volume613
    ISSN (Print)0076-6879
    ISSN (Electronic)1557-7988

    Keywords

    • Crystallization
    • Membrane protein complexes
    • Oxygen evolution
    • Photosystem II
    • Purification
    • Water oxidation

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology

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