Tonoplast-enriched vesicles were prepared from suspension-cultured Populus euphratica Oliv. cells by differential centrifugation and discontinuous sucrose density gradient centrifugation. The properties of the proton ...Tonoplast-enriched vesicles were prepared from suspension-cultured Populus euphratica Oliv. cells by differential centrifugation and discontinuous sucrose density gradient centrifugation. The properties of the proton pumping activity of H+-ATPases in tonoplast vesicles were studied by acridine orange fluorescent quenching measured at 22 degreesC. The proton pumping activity of ATPase was ATP-dependent with apparent Michaelis-Menten Constant (K-m) for ATP about 0.65 mmol/L. The optimal pH for H+-ATPases activity was 7.5. The proton pumping activity of H+-ATPase could be initiated by some divalent cations, Mg2+ being highly efficient, much more than Fe2+; and Ca2+, Cu2+ and Zn2+ were inefficient under the experimental condition. The proton translocation could be stimulated by halide anions, with potencies decreasing in the order Cl- > Br- > I- > F-. The proton pumping activity was greatly inhibited by N-ethylmaleimide (NEM), N, N'-dicyclohexylcarbodiimide (DCCD), NO3- and Bafilomycin A(1), but not by orthovanadate and azide. These results demonstrated that the H+-ATPase in the tonoplast of Populus euphratica belonged to vacuolar type ATPase. This work was the first time that tonoplast-enriched vesicles were isolated from Populus euphratica cells.展开更多
The combination of needling technique of Mind Regulation with Taoism Concentration is to promote the propagated sensation along the channels (PSC) and the needling sensation to the affected area in order to activate...The combination of needling technique of Mind Regulation with Taoism Concentration is to promote the propagated sensation along the channels (PSC) and the needling sensation to the affected area in order to activate the potential regulation of human body. Prof. Yang Yun-kuan used this combination to treat some pain diseases, nervous system's diseases and intractable chronic diseases and got a better result than the general techniques, This is the introduction to the manipulation of this technique and its clinical application.展开更多
文摘Tonoplast-enriched vesicles were prepared from suspension-cultured Populus euphratica Oliv. cells by differential centrifugation and discontinuous sucrose density gradient centrifugation. The properties of the proton pumping activity of H+-ATPases in tonoplast vesicles were studied by acridine orange fluorescent quenching measured at 22 degreesC. The proton pumping activity of ATPase was ATP-dependent with apparent Michaelis-Menten Constant (K-m) for ATP about 0.65 mmol/L. The optimal pH for H+-ATPases activity was 7.5. The proton pumping activity of H+-ATPase could be initiated by some divalent cations, Mg2+ being highly efficient, much more than Fe2+; and Ca2+, Cu2+ and Zn2+ were inefficient under the experimental condition. The proton translocation could be stimulated by halide anions, with potencies decreasing in the order Cl- > Br- > I- > F-. The proton pumping activity was greatly inhibited by N-ethylmaleimide (NEM), N, N'-dicyclohexylcarbodiimide (DCCD), NO3- and Bafilomycin A(1), but not by orthovanadate and azide. These results demonstrated that the H+-ATPase in the tonoplast of Populus euphratica belonged to vacuolar type ATPase. This work was the first time that tonoplast-enriched vesicles were isolated from Populus euphratica cells.
文摘The combination of needling technique of Mind Regulation with Taoism Concentration is to promote the propagated sensation along the channels (PSC) and the needling sensation to the affected area in order to activate the potential regulation of human body. Prof. Yang Yun-kuan used this combination to treat some pain diseases, nervous system's diseases and intractable chronic diseases and got a better result than the general techniques, This is the introduction to the manipulation of this technique and its clinical application.