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Complex formation in plant thylakoid membranes. Competition studies on membrane protein interactions using synthetic peptide fragments

Abstract

Thylakoid membranes of pea were used to study competition between extra-membrane fragments and their parental membrane-bound proteins. Phosphorylated and unphosphorylated fragments of light harvesting complex II (LHC II) from higher plants were used to compete with LHC II for interactions with itself and with other thylakoid protein complexes. Effects of these peptide fragments of LHC II and of control peptides were followed by 80 K chlorophyll fluorescence spectroscopy of isolated thylakoids. The phosphorylated LHC II fragment competes with membrane-bound phosphoproteins in the phosphatase reaction. The same fragment accelerates the process of dark-to-light adaptation and decreases the rate of the light-to-dark adaptation when these are followed by fluorescence spectroscopy. In contrast, the non-phosphorylated LHC II peptide does not affect the rate of adaptation but produces results consistent with inhibition of formation of a quenching complex. In this quenching complex we propose that LHC II remains inaccessible to the LHC II kinase, explaining an observed decrease in LHC II phosphorylation in the later stages of the time-course of phosphorylation. The most conspicuous protein which is steadily phosphorylated during the time-course of phosphorylation is the 9 kDa (psbH) protein. The participation of the phosphorylated form of psbH in the quenching complex, where it is inaccessible to the phosphatase, may explain its anomalously slow dephosphorylation. The significance of the proposed complex of LHC II with phospho-psbH is discussed.

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Abbreviations

LHC II:

light harvesting complex II

PS II:

Photosystem II

PS I:

Photosystem I

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Stys, D., Stancek, M., Cheng, L. et al. Complex formation in plant thylakoid membranes. Competition studies on membrane protein interactions using synthetic peptide fragments. Photosynth Res 44, 277–285 (1995). https://doi.org/10.1007/BF00048601

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  • DOI: https://doi.org/10.1007/BF00048601

Key words

  • light harvesting complex II
  • membrane proteins
  • macromolecular recognition
  • protein phosphorylation
  • excitation energy transfer
  • energy quenching
  • psb H