The 28 kDa apoprotein of CP 26 in PS II binds copper

Abstract

Photosystem II (PS II) particles isolated from spinach in the presence of 10 μM CuSO4 contained 1.2 copper/300 Chl that was resistant to EDTA. When CuSO4 was not added during the isolation, PS II particles contained variable amounts of copper resistant to EDTA (0.1-1.1 copper/300 Chl). No correlation was found between copper content and oxygen evolving capacity of the PS II particles. To identify the copper binding protein, we developed a fractionation procedure which included solubilisation of PS II particles followed by precipitation with polyethylene glycol. A 22-fold purification of copper with respect to protein was achieved for a 28 kDa protein. Partial amino acid sequence of a 13 kDa fragment, obtained after V8 (endo Glu-C) protease treatment, showed identity with CP 26 over a 14 amino acid stretch. EPR measurements on the purified protein suggest oxygen and/or nitrogen as ligands for copper but tend to exclude sulfur. We conclude that the 28 kDa apoprotein of CP 26 from spinach binds one copper per molecule of CP 26. A possible function for this copper protein in the xanthophyll cycle is discussed.