One-step isolation and biochemical characterization of a highly active plant PSII monomeric core

Photosynth Res. 2011 May;108(1):33-46. doi: 10.1007/s11120-011-9650-4. Epub 2011 Apr 13.

Abstract

We describe a one-step detergent solubilization protocol for isolating a highly active form of Photosystem II (PSII) from Pisum sativum L. Detailed characterization of the preparation showed that the complex was a monomer having no light harvesting proteins attached. This core reaction centre complex had, however, a range of low molecular mass intrinsic proteins as well as the chlorophyll binding proteins CP43 and CP47 and the reaction centre proteins D1 and D2. Of particular note was the presence of a stoichiometric level of PsbW, a low molecular weight protein not present in PSII of cyanobacteria. Despite the high oxygen evolution rate, the core complex did not retain the PsbQ extrinsic protein although there was close to a full complement of PsbO and PsbR and partial level of PsbP. However, reconstitution of PsbP and PsbPQ was possible. The presence of PsbP in absence of LHCII and other chlorophyll a/b binding proteins confirms that LHCII proteins are not a strict requirement for the assembly of this extrinsic polypeptide to the PSII core in contrast with the conclusion of Caffarri et al. (2009).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Blotting, Western
  • Electrophoresis, Polyacrylamide Gel
  • Membrane Proteins / chemistry
  • Membrane Proteins / isolation & purification*
  • Membrane Proteins / metabolism
  • Microscopy, Electron, Transmission
  • Oxygen / analysis
  • Oxygen / metabolism
  • Photosystem II Protein Complex / chemistry*
  • Photosystem II Protein Complex / isolation & purification*
  • Photosystem II Protein Complex / metabolism
  • Pisum sativum / chemistry*
  • Pisum sativum / metabolism
  • Plant Proteins / chemistry
  • Plant Proteins / isolation & purification*
  • Plant Proteins / metabolism
  • Protein Subunits / chemistry
  • Protein Subunits / isolation & purification
  • Protein Subunits / metabolism
  • Spectrometry, Mass, Electrospray Ionization
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Tandem Mass Spectrometry
  • Temperature
  • Thylakoids / chemistry*
  • Thylakoids / metabolism

Substances

  • Membrane Proteins
  • Photosystem II Protein Complex
  • Plant Proteins
  • Protein Subunits
  • Oxygen