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Scientific article
English

Isolation of a novel heterodimeric PSII complex via strep-tagged PsbO

Published inBiochimica et biophysica acta. Bioenergetics, vol. 1864, no. 2, 148953
Publication date2023-04
Abstract

The multi-subunit membrane protein complex photosystem II (PSII) catalyzes the light-driven oxidation of water and with this the initial step of photosynthetic electron transport in plants, algae, and cyanobacteria. Its biogenesis is coordinated by a network of auxiliary proteins that facilitate the stepwise assembly of individual subunits and cofactors, forming various intermediate complexes until fully functional mature PSII is present at the end of the process. In the current study, we purified PSII complexes from a mutant line of the thermophilic cyanobacterium Thermosynechococcus vestitus BP-1 in which the extrinsic subunit PsbO, characteristic for active PSII, was fused with an N-terminal Twin-Strep-tag. Three distinct PSII complexes were separated by ion- exchange chromatography after the initial affinity purification. Two complexes differ in their oligomeric state (monomeric and dimeric) but share the typical subunit composition of mature PSII. They are characterized by the very high oxygen evolving activity of approx. 6000 μmol O2⋅(mg Chl⋅h) 1. Analysis of the third (heterodimeric) PSII complex revealed lower oxygen evolving activity of approx. 3000 μmol O2⋅(mg Chl⋅h) 1 and a manganese content of 2.7 (±0.2) per reaction center compared to 3.7 (±0.2) of fully active PSII. Mass spectrometry and time-resolved fluorescence spectroscopy further indicated that PsbO is partially replaced by Psb27 in this PSII fraction, thus implying a role of this complex in PSII repair.

eng
Keywords
  • Heterodimer
  • Photosynthesis
  • Photosystem II
  • Thermosynechococcus vestitus BP-1
  • Time resolved fluorescence spectroscopy
  • Twin-Strep-tag
Citation (ISO format)
LAMBERTZ, Jan et al. Isolation of a novel heterodimeric PSII complex via strep-tagged PsbO. In: Biochimica et biophysica acta. Bioenergetics, 2023, vol. 1864, n° 2, p. 148953. doi: 10.1016/j.bbabio.2022.148953
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ISSN of the journal0005-2728
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