Purification of fully active and crystallizable photosystem II from thermophilic cyanobacteria

Keisuke Kawakami, Jian-Ren Shen

Research output: Chapter in Book/Report/Conference proceedingChapter

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 - Jan 1 2018

Publication series

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

Fingerprint

Photosystem II Protein Complex
Cyanobacteria
Purification
Dimers
Native Polyacrylamide Gel Electrophoresis
Oxygen
Light
Photosynthesis
Water
Molecular oxygen
Molecular mass
Crystallization
Chromatography
Atmosphere
Gel Chromatography
Polyacrylamide Gel Electrophoresis
Absorption spectra
Membrane Proteins
Diffraction
Monomers

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Kawakami, K., & Shen, J-R. (2018). Purification of fully active and crystallizable photosystem II from thermophilic cyanobacteria. In F. Armstrong (Ed.), Methods in Enzymology (pp. 1-16). (Methods in Enzymology; Vol. 613). Academic Press Inc.. https://doi.org/10.1016/bs.mie.2018.10.002

Purification of fully active and crystallizable photosystem II from thermophilic cyanobacteria. / Kawakami, Keisuke; Shen, Jian-Ren.

Methods in Enzymology. ed. / Fraser Armstrong. Academic Press Inc., 2018. p. 1-16 (Methods in Enzymology; Vol. 613).

Research output: Chapter in Book/Report/Conference proceedingChapter

Kawakami, K & Shen, J-R 2018, Purification of fully active and crystallizable photosystem II from thermophilic cyanobacteria. in F Armstrong (ed.), Methods in Enzymology. Methods in Enzymology, vol. 613, Academic Press Inc., pp. 1-16. https://doi.org/10.1016/bs.mie.2018.10.002
Kawakami K, Shen J-R. Purification of fully active and crystallizable photosystem II from thermophilic cyanobacteria. In Armstrong F, editor, Methods in Enzymology. Academic Press Inc. 2018. p. 1-16. (Methods in Enzymology). https://doi.org/10.1016/bs.mie.2018.10.002
Kawakami, Keisuke ; Shen, Jian-Ren. / Purification of fully active and crystallizable photosystem II from thermophilic cyanobacteria. Methods in Enzymology. editor / Fraser Armstrong. Academic Press Inc., 2018. pp. 1-16 (Methods in Enzymology).
@inbook{d9952fc667fb45478c59bcdea94506f6,
title = "Purification of fully active and crystallizable photosystem II from thermophilic cyanobacteria",
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.",
keywords = "Crystallization, Membrane protein complexes, Oxygen evolution, Photosystem II, Purification, Water oxidation",
author = "Keisuke Kawakami and Jian-Ren Shen",
year = "2018",
month = "1",
day = "1",
doi = "10.1016/bs.mie.2018.10.002",
language = "English",
isbn = "9780128163610",
series = "Methods in Enzymology",
publisher = "Academic Press Inc.",
pages = "1--16",
editor = "Fraser Armstrong",
booktitle = "Methods in Enzymology",
address = "United States",

}

TY - CHAP

T1 - Purification of fully active and crystallizable photosystem II from thermophilic cyanobacteria

AU - Kawakami, Keisuke

AU - Shen, Jian-Ren

PY - 2018/1/1

Y1 - 2018/1/1

N2 - 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.

AB - 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.

KW - Crystallization

KW - Membrane protein complexes

KW - Oxygen evolution

KW - Photosystem II

KW - Purification

KW - Water oxidation

UR - http://www.scopus.com/inward/record.url?scp=85057060716&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85057060716&partnerID=8YFLogxK

U2 - 10.1016/bs.mie.2018.10.002

DO - 10.1016/bs.mie.2018.10.002

M3 - Chapter

C2 - 30509462

AN - SCOPUS:85057060716

SN - 9780128163610

T3 - Methods in Enzymology

SP - 1

EP - 16

BT - Methods in Enzymology

A2 - Armstrong, Fraser

PB - Academic Press Inc.

ER -