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Photosystem II (PSII) splits water and drives electron transfer to plastoquinone via photochemical reactions using light energy. It is surrounded by light‐harvesting complex II (LHCII) to form the PSII–LHCII supercomplex. Complete characterization of its structure and function has, however, been hampered due to instability of the complex in the presence of detergent. To overcome this problem, we developed a new procedure for purifying the PSII–LHCII supercomplexes of Chlamydomonas reinhardtii employing amphipol A8‐35. The obtained supercomplexes showed little LHCII dissociation even 4 days after purification. Oxygen‐evolving activity was retained within amphipol if the extrinsic polypeptides were kept associated by betaine. Electron microscopy revealed that this method also improved structural uniformity and that the major organization was C2S2M2L2.

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