Structural, Biochemical and Biophysical Characterization of Four Oxygen-Evolving Photosystem II Preparations From Spinach

Terri Goodman Dunahay, L. Andrew Staehelin, Michael Seibert, Paula D. Ogilvie, Steven P. Berg

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205 Scopus Citations


Four procedures utilizing different detergent and salt conditions were used to isolate oxygen-evolving Photosystem II (PS II) preparations from spinach thylakoid membranes. These PS II preparations have been characterized by freeze-fracture electron microscopy, SDS-polyacrylamide gel electrophoresis, steady-state and pulsed oxygen evolution, 77 K fluorescence, and room-temperature electron paramagnetic resonance. All of the O2-evolving PS II samples were found to be highly purified grana membrane fractions composed of paired, appressed membrane fragments. The lumenal surfaces of the membranes and thus the O2-evolving enzyme complex, are directly exposed to the external environment. Biochemical and biophysical analyses indicated that all four preparations are enriched in the chlorophyll a b-light-harvesting complex and Photosystem II, and depleted to varying degrees in the stroma-associated components, Photosystem I and the CF1-ATPase. The four PS II samples also varied in their cytochrome f content. All preparations showed enhanced stability of oxygen production and oxygen-rate electrode activity compared to control thylakoids, apparently promoted by low concentrations of residual detergent in the PS II preparations. A model is presented which summarizes the effects of the salt and detergent treatments on thylakoid structure and, consequently, on the configuration and composition of the oxygen-evolving PS II samples.

Original languageAmerican English
Pages (from-to)179-193
Number of pages15
JournalBBA - Bioenergetics
Issue number2
StatePublished - 1984
Externally publishedYes

NREL Publication Number

  • ACNR/JA-233-4744


  • (Spinach chloroplast)
  • ESR
  • Fluorescence
  • Freeze-fracture
  • Oxygen evolution
  • Photosystem II


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