On the Reconstitution of Photophosphorylation in Chloroplast Membranes

Varda Shoshan, Noun Shavit

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A high degree of reconstitution of photophosphorylation was achieved using resolved chloroplast particles obtained by incubation with 1 mM EDTA at relatively high chlorophyll concentrations (about 400 μg/ml). The resolved particles lost about 95% and 75% of their phosphorylative and proton transport activity respectively. Upon reconstitution with either crude or purified chloroplast coupling factor preparations, almost full activity was recovered. Control of electron transport by phosphorylating agents was also observed in these reconstituted preparations. Highly reconstituted partieles were also obtained by incubation of chloroplasts, at low chlorophyll concentrations, with EDTA up to 10 mM. Above this concentration of EDTA, poor uncoupling was observed. Gel electrophoresis of supernatant fractions obtained from incubations with EDTA under conditions which produced resolved particles with a poor degree of reconstitution revealed a protein fraction not appearing in supernatants from highly reconstituted particles. The degree of reconstitution was inversely correlated with the appearance of this protein band.

Original languageEnglish
Pages (from-to)355-360
JournalEuropean Journal of Biochemistry
Issue number2
StatePublished - 1 Jan 1973


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