TAILIEUCHUNG - Báo cáo khoa học: Reconstitution of coupled fumarate respiration in liposomes by incorporating the electron transport enzymes isolated from Wolinella succinogenes

Hydrogenase and fumarate reductase isolated fromWoli-nella succinogeneswere incorporated into liposomes con-taining menaquinone. The two enzymes were found to be oriented solely to the outside of the resulting proteolipo-somes. The proteoliposomes catalyzed fumarate reduction by H2 which generated an electrical proton potential (Dw V, negative inside) in the same direction as that gen-erated by fumarate respiration in cells ofW. succinogenes. | Eur. J. Biochem. 269 1974-1983 2002 FEBS 2002 doi Reconstitution of coupled fumarate respiration in liposomes by incorporating the electron transport enzymes isolated from Wolinella succinogenes Simone Biel1. Jora Simon1. Roland Gross1 Teresa Ruiz2 Maarten Ruitenbera3 and Achim Kroaer1 1Institut fur Mikrobiologie Johann Wolfgang Goethe-Universitat Frankfurt am Main Germany 2Max-Planck-Institut fur Biophysik Abteilung Strukturbiologie Frankfurt am Main Germany 3Max-Planck-Institut fur Biophysik Abteilung Biophysikalische Chemie Frankfurt am Main Germany Hydrogenase and fumarate reductase isolated from Woli-nella succinogenes were incorporated into liposomes containing menaquinone. The two enzymes were found to be oriented solely to the outside of the resulting proteolipo-somes. The proteoliposomes catalyzed fumarate reduction by H2 which generated an electrical proton potential AW V negative inside in the same direction as that generated by fumarate respiration in cells of W. succinogenes. The H e ratio brought about by fumarate reduction with H2 in proteoliposomes in the presence of valinomycin and external K was approximately 1. The same AW and H e ratio was associated with the reduction of 2 3-dimethyl-1 4-naphthoquinone DMN by H2 in proteoliposomes containing menaquinone and hydrogenase with or without fumarate reductase. Proteoliposomes containing menaqui-none and fumarate reductase with or without hydrogenase catalyzed fumarate reduction by DMNH2 which did not generate a AW. Incorporation of formate dehydrogenase together with fumarate reductase and menaquinone resulted in proteoliposomes catalyzing the reduction of fumarate or DMN by formate. Both reactions generated a AW of V negative inside . The H e ratio of formate oxidation by menaquinone or DMN was close to 1. The results demonstrate for the first time that coupled fumarate respiration can be restored in liposomes using the well characterized electron transport

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