The contribution of two components (ΔpH and ΔE) of the proton motive force to photosynthesis of C. reinhardtii was studied. Valinomycin, a photophosphorylation uncoupler, decreased significantly the fast phase (related mainly to the membrane electric potential) of millisecond delayed light emission (ms-DLE) of C. reinhardtii. Nigericin, another photophosphorylation uncoupler, decreased the slow phase (related mainly to the proton gradient) and partly also the fast phase of ms-DLE. Both valinomycin and nigericin decreased the net ATP content and photosynthetic rate of C. reinhardtii, but the inhibition by nigericin was stronger than that by valinomycin. Hence both components of the proton motive force contribute to photosynthesis and although the contribution of ΔpH is larger than that of ΔE, the latter is not negligible in photosynthesis of C. reinhardtii. and Qing-Xiu Tang, Jia-Mian Wei.
In the mutant CC-1047 of Chlamydomonas reinhardtii, LDS-PAGE showed that the chlorophyll-protein complex I (CPI) is almost absent. The mutant could not grow in a culture medium without organic carbon source while the wild type (WT) C. reinhardtii grew quickly. When an organic carbon source was added into the culture medium, the mutant grew almost as well as WT. The rate of photosystem 1 (PS1) electron transport (DCPIP→MV) and the rate of whole chain electron transport (H2O→MV) of chloroplasts of the CC-1047 mutant were both lower than those of WT. The photophosphorylation activity, photosynthetic O2 evolution rate, and rate of NADP+ photoreduction of CC-1047 were also much lower than the activities of WT. There were some differences in ATPase activity between the mutant and WT. Two different activation ways were used to activate the latent ATPase using methanol and dithiothreitol (DTT) as activation substrate. More methanol and DTT were required for the mutant than WT to obtain the maximum activity. Thus the photosynthetic apparatus could not operate normally when CPI was absent because of the abnormal PS1 electron transport. Meanwhile, the other adjacent complexes of the thylakoid membrane, for example, ATP synthase complex, were slightly affected. and Qing-Xiu Tang, Zhang-Lin Ni, Jia-Mian Wei.
The conserved residue Thr42 of ε-subunit of the chloroplast ATP synthase of maize (Zea mays L.) was substituted with Cys, Arg, and Ile, respectively, through site-directed mutagenesis. The over-expressed and refolded ε-proteins were purified by chromatography on DEAE-cellulose and FPLC on mono-Q column, which were as biologically active (inhibiting Ca2+-ATPase activity and blocking proton gate) as the native ε subunit isolated from chloroplasts. The εT42C and εT42R showed higher inhibitory activities on the soluble CF1(-ε) Ca2+-ATPase than the εWT. The εT42I inhibited the Ca2+-ATPase activity of soluble CF1 and restored photophosphorylation activity of membrane-bound CF1 deficient in ε the most efficiently. Far-ultraviolet CD spectra showed that the portions of α-helix and β-sheet structures of the three mutants were somewhat different from εWT. Thus the conserved residue Thr42 may be important for maintaining the structure and function of the ε-subunit and the basic functions of the ε-subunit as far as an inhibitor of Ca2+-ATPase and the proton gate are related. and Zhang-Lin Ni, Da-Fu Wang, Jia-Mian Wei.