caa a22 4500 445890053 CHVBK 20180317145613.0 cr unu---uuuuu 170323e20110501xx s 000 0 eng 10.1007/s11120-011-9650-4 doi (NATIONALLICENCE)springer-10.1007/s11120-011-9650-4 One-step isolation and biochemical characterization of a highly active plant PSII monomeric core [Elektronische Daten] [Cristina Pagliano, Fabiana Chimirri, Guido Saracco, Francesco Marsano, James Barber] 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). Springer Science+Business Media B.V., 2011 Photosystem II nationallicence Monomer nationallicence Pisum sativum nationallicence PsbP nationallicence Oxygen evolving activity nationallicence BN-PAGE : Blue native polyacrylamide gel electrophoresis nationallicence Chl : Chlorophyll nationallicence DCBQ : 2,6-Dichlorobenzoquinone nationallicence DCMU : 3-(3,4-Dichloropheny1)-l,l-dimethylurea nationallicence EM : Electron microscopy nationallicence ESI : Electrospray ionization nationallicence K3Fe(CN)6 : Potassium ferricyanide nationallicence LC : Liquid chromatography nationallicence LHC : Light harvesting complex nationallicence MALDI-TOF : Matrix-assisted laser desorption/ionization-time of flight nationallicence MS : Mass spectrometry nationallicence OEC : Oxygen evolving complex nationallicence PEG6000 : Polyethylene glycol Mr 6000 nationallicence PSII : Photosystem II nationallicence SDS-PAGE : Sodium dodecyl sulphate polyacrylamide gel electrophoresis nationallicence α-DM : alpha-Dodecylmaltoside nationallicence β-DM : beta-Dodecylmaltoside nationallicence Pagliano Cristina Department of Materials Science and Chemical Engineering - BioSolar Lab, Politecnico di Torino, Viale T. Michel 5, 15121, Alessandria, Italy aut Chimirri Fabiana Department of Materials Science and Chemical Engineering - BioSolar Lab, Politecnico di Torino, Viale T. Michel 5, 15121, Alessandria, Italy aut Saracco Guido Department of Materials Science and Chemical Engineering - BioSolar Lab, Politecnico di Torino, Viale T. Michel 5, 15121, Alessandria, Italy aut Marsano Francesco Department of Environmental and Life Sciences-Proteomic Facility, University of Piemonte Orientale, Viale T. Michel 11, 15121, Alessandria, Italy aut Barber James Department of Materials Science and Chemical Engineering - BioSolar Lab, Politecnico di Torino, Viale T. Michel 5, 15121, Alessandria, Italy aut Photosynthesis Research Springer Netherlands 108/1(2011-05-01), 33-46 0166-8595 108:1<33 2011 108 11120 https://doi.org/10.1007/s11120-011-9650-4 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11120-011-9650-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Pagliano Cristina Department of Materials Science and Chemical Engineering - BioSolar Lab, Politecnico di Torino, Viale T. Michel 5, 15121, Alessandria, Italy aut NATIONALLICENCE 700 1- Chimirri Fabiana Department of Materials Science and Chemical Engineering - BioSolar Lab, Politecnico di Torino, Viale T. Michel 5, 15121, Alessandria, Italy aut NATIONALLICENCE 700 1- Saracco Guido Department of Materials Science and Chemical Engineering - BioSolar Lab, Politecnico di Torino, Viale T. Michel 5, 15121, Alessandria, Italy aut NATIONALLICENCE 700 1- Marsano Francesco Department of Environmental and Life Sciences-Proteomic Facility, University of Piemonte Orientale, Viale T. Michel 11, 15121, Alessandria, Italy aut NATIONALLICENCE 700 1- Barber James Department of Materials Science and Chemical Engineering - BioSolar Lab, Politecnico di Torino, Viale T. Michel 5, 15121, Alessandria, Italy aut NATIONALLICENCE 773 0- Photosynthesis Research Springer Netherlands 108/1(2011-05-01), 33-46 0166-8595 108:1<33 2011 108 11120 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer