Identification of thylakoid membrane thermal transitions in Synechocystis sp. PCC6803 photosynthetic mutants

Abstract

We used differential scanning calorimetry (DSC) as a technique capable of identifying photosynthetic complexes on the basis of their calorimetric transitions. Annotation of thermal transitions was carried out with thylakoid membranes isolated from various photosynthetic mutants of Synechocystis sp. PCC6803. The thylakoid membranes exhibited seven major DSC bands between 40 and 85°C. The heat sorption curves were analyzed both by mathematical deconvolution of the overall endotherms and by a subsequent annealing procedure. The successive annealing procedure proved to be more reliable technique than mathematical deconvolution in assigning thermal transitions. The main DSC band, around 47°C, resulting from the high enthalpy change that corresponds to non-interacting complex of PSII, was assigned using the PSI-less/apcE ⁻ mutant cells. Another band around 68–70°C relates to the denaturation of PSII surrounded by other proteins of the photosynthetic complexes in wild type and PSI-less/apcE ⁻ cells. A further major transition found at 82–84°C corresponds to the PSI core complex of wild type and PSII-deficient BE cells. Other transition bands between 50–67 and 65–75°C are believed to relate to ATP synthase and cytochrome b ₆ f, respectively. These thermal transitions were obtained with thylakoids isolated from PSI⁻/PSII⁻ mutant cells. Some minor bands determined at 59 and 83–84°C correspond to an unknown complex and NADH dehydrogenase, respectively. These annotations were done by PSI-less/apcE ⁻ and PSI⁻/PSII⁻ mutants.