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PreprintCC BYbioRxiv

Mass spectrometric identification of a PSI-LHCI-cytochrome b6f complex in both Arabidopsis stn7 and pph1

  1. 1University of Stavanger

Abstract

Native PAGE separation of digitonin-solubilized thylakoid stroma lamellae resolves PSI and cytochrome b6f into separate bands. Mass spectrometric analysis of the 1D native gel bands showed that the bands are baskets of co-migrating complexes along the molecular size axis. Hierarchical clustering confirmed the separation of dimeric cytochrome b6f from PSI in 1D LN-PAGE. Interval-zone free-flow electrophoresis separated the same extracts by charge in a flowing buffer film, and the continuous fractionation indicated co-migration of complexes without the in-gel structural hindrances. A second-dimension LN-PAGE separated the complexes co-migrating in iZE into complexes with defined molecular size, allowing in-gel mass spectrometry identification in the gel bands. Both genotypes showed a 720 kDa fluorescent band in which cytochrome f (PetA) and cytochrome b6 (PetB) join the PSI-LHCI node (stn7, d = 0.50; pph1, d = 0.34). Lhcb1 and Lhcb2 are absent from the 720 kDa node. The 753 kDa band is specific to pph1 (d = 0.38). Phosphorylated LHCII1,2 joins PSI-LHCI there, and PetB is detected in every slice of that band without becoming a leaf of the tree.

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BibTeX
@misc{eichacker2026mass,
  title = {{Mass spectrometric identification of a PSI-LHCI-cytochrome b6f complex in both Arabidopsis stn7 and pph1}},
  author = {L. A. Eichacker and J. Arnold and G. Fucile and A. K. Vatland and K. Stensen and G. Weber},
  howpublished = {bioRxiv},
  year = {2026},
  month = oct,
  doi = {10.64898/2026.10.16.759642},
  url = {https://www.biorxiv.org/content/10.64898/2026.10.16.759642v1},
  note = {CC BY},
}
RIS
TY  - UNPB
TI  - Mass spectrometric identification of a PSI-LHCI-cytochrome b6f complex in both Arabidopsis stn7 and pph1
AU  - L. A. Eichacker
AU  - J. Arnold
AU  - G. Fucile
AU  - A. K. Vatland
AU  - K. Stensen
AU  - G. Weber
PY  - 2026
DA  - 2026/10/16
PB  - bioRxiv
DO  - 10.64898/2026.10.16.759642
UR  - https://www.biorxiv.org/content/10.64898/2026.10.16.759642v1
AB  - Native PAGE separation of digitonin-solubilized thylakoid stroma lamellae resolves PSI and cytochrome b6f into separate bands. Mass spectrometric analysis of the 1D native gel bands showed that the bands are baskets of co-migrating complexes along the molecular size axis. Hierarchical clustering confirmed the separation of dimeric cytochrome b6f from PSI in 1D LN-PAGE. Interval-zone free-flow electrophoresis separated the same extracts by charge in a flowing buffer film, and the continuous fractionation indicated co-migration of complexes without the in-gel structural hindrances. A second-dimension LN-PAGE separated the complexes co-migrating in iZE into complexes with defined molecular size, allowing in-gel mass spectrometry identification in the gel bands. Both genotypes showed a 720 kDa fluorescent band in which cytochrome f (PetA) and cytochrome b6 (PetB) join the PSI-LHCI node (stn7, d = 0.50; pph1, d = 0.34). Lhcb1 and Lhcb2 are absent from the 720 kDa node. The 753 kDa band is specific to pph1 (d = 0.38). Phosphorylated LHCII1,2 joins PSI-LHCI there, and PetB is detected in every slice of that band without becoming a leaf of the tree.
ER  -