A strain of Synechocystis sp. PCC 6803 expressing the yellow fluorescent protein (YFP) fused to the C-terminus of the PsaF subunit of PSI has been constructed and used to isolate native PSI complexes employing the GFP-Trap®, an efficient immunoprecipitation system which recognizes the green fluorescent protein (GFP) and its variants. The protein analysis and spectroscopic characterization of the preparation revealed an isolate of trimeric and monomeric PSI complexes, which showed minimal unspecific contamination as demonstrated by comparison with the wild type control. Interestingly, we detected CP43 subunits of PSII and small amounts of PSII core complexes specifically pulled-down with the YFP-PSI, supporting the association of PSII assembly modules and intermediate assembly complexes with PSI, as observed in our previous studies. The results demonstrate that the GFP-Trap® system represents an excellent tool for studies of PSI biogenesis and interconnection of PSI and PSII assembly processes., A. Strašková, J. Knoppová, J. Komenda., and Obsahuje bibliografické odkazy
Our study examined the relationship between photosynthetic performance and activities of key photosynthetic enzymes to understand the photosynthetic variation and reasons for the variation during dormancy induction under different photoperiods in peach (Prunus persica L. cv. Chunjie). Furthermore, the study explained the changes in the key enzymes from the viewpoint of differential proteomics. The results showed that the leaf net photosynthetic rate (PN) and stomatal conductance tended to decrease, while the intercellular CO2 concentration rose, which indicated that the reduced PN resulted from nonstomatal limitation. During the dormancy induction period, the activities of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and phosphoenolpyruvate carboxylase (PEPC) declined, which was the main reason for the reduced PN. Two-dimensional electrophoresis maps and differential protein identification demonstrated that the decrease in activity of the photosynthetic enzymes was mainly due to enzymatic degradation. The enzyme degradation by a long-day treatment occurred later and to a lesser degree than that of the short-day treatment. In the long-day treatment, the carboxylation activity of Rubisco was higher than that of the control treatment, and the PEPC activity and the ratio of the PEPC/Rubisco activity were lower than the corresponding activities during the control treatment. These differences under long-day conditions were significant but did not occur in the short-day treatment, suggesting that the C4 pathway might be more active under short-day conditions., H.-S. Zhang, D.-M. Li, Q.-P. Tan, H.-Y. Gao, D.-S. Gao., and Obsahuje bibliografii