論文

査読有り
2010年1月

Purification and characterization of photosystem I complex from Synechocystis sp PCC 6803 by expressing histidine-tagged subunits

BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS
  • Hisako Kubota
  • ,
  • Isamu Sakurai
  • ,
  • Kenta Katayama
  • ,
  • Naoki Mizusawa
  • ,
  • Shunsuke Ohashi
  • ,
  • Masami Kobayashi
  • ,
  • Pengpeng Zhang
  • ,
  • Eva-Mari Aro
  • ,
  • Hajime Wada

1797
1
開始ページ
98
終了ページ
105
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1016/j.bbabio.2009.09.001
出版者・発行元
ELSEVIER SCIENCE BV

We generated Synechocystis sp. PCC 6803 strains, designated F-His and J-His, which express histidine-tagged PsaF and PsaJ subunits, respectively, for simple purification of the photosystem I (PSI) complex. Six histidine residues were genetically added to the C-terminus of the PsaF subunit in F-His cells and the N-terminus of the PsaJ subunit in J-His cells. The histidine residues introduced had no apparent effect on photoautotrophic growth of the cells or the activity of PSI and PSII in thylakoid membranes. PSI complexes could be simply purified from the F-His and J-His cells by Ni2+-affinity column chromatography. When thylakoid membranes corresponding to 20 mg chlorophyll were used, PSI complexes corresponding to about 7 mg chlorophyll could be purified in both strains. The purified PSI complexes could be separated into monomers and trimers by ultracentrifugation in glycerol density gradient and high activity was recorded for trimers isolated from the F-His and J-His strains. Blue-Native PAGE and SDS-PAGE analysis of monomers and trimers indicated the existence of two distinct monomers with different subunit compositions and no contamination of PSI with other complexes, such as PSII and Cyt b(6)f. Further analysis of proteins and lipids in the purified PSI indicated the presence of novel proteins in the monomers and about six lipid molecules per monomer unit in the trimers. These results demonstrate that active PSI complexes can be simply purified from the constructed strains and the strains are very useful tools for analysis of PSI. (C) 2009 Elsevier B.V. All rights reserved.

Web of Science ® 被引用回数 : 46

リンク情報
DOI
https://doi.org/10.1016/j.bbabio.2009.09.001
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000272856900012&DestApp=WOS_CPL
ID情報
  • DOI : 10.1016/j.bbabio.2009.09.001
  • ISSN : 0005-2728
  • eISSN : 0006-3002
  • Web of Science ID : WOS:000272856900012

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