TAILIEUCHUNG - Báo cáo khoa học: Insights into the design of a hybrid system between Anabaena ferredoxin-NADP+ reductase and bovine adrenodoxin

The opportunity to design enzymatic systems is becoming more feasible due to detailed knowledge of the structure of many proteins. As a first step, investigations have aimed to redesign already existing systems, so that they can performa function different from the one for which they were syn-thesized. We have investigated the interaction of electron transfer proteins fromdifferent systems inorder to check the possibility of heterologous reconstitution among members of different chains. | Eur. J. Biochem. 270 726-735 2003 FEBS 2003 doi Insights into the design of a hybrid system between Anabaena ferredoxin-NADP reductase and bovine adrenodoxin Merche Faro1 Burkhard Schiffler2 Achim Heinz2 Isabel Nogues1 Milagros Medina1 Rita Bernhardt2 and Carlos Gomez-Moreno1 1 Departamento de Bioquimica y Biologia Molecular y Celular Facultad de Ciencias Universidad de Zaragoza Spain 2Biochemie Universitat des Saarlandes Saarbrucken Germany The opportunity to design enzymatic systems is becoming more feasible due to detailed knowledge of the structure of many proteins. As a first step investigations have aimed to redesign already existing systems so that they can perform a function different from the one for which they were synthesized. We have investigated the interaction of electron transfer proteins from different systems in order to check the possibility of heterologous reconstitution among members of different chains. Here it is shown that ferredoxin-NADP reductase from Anabaena and adrenodoxin from bovine adrenal glands are able to form optimal complexes for thermodynamically favoured electron transfer reactions. Thus electron transfer from ferredoxin-NADP reductase to adrenodoxin seems to proceed through the formation of at least two different complexes whereas electron transfer from adrenodoxin to ferredoxin-NADP reductase does not take place due because it is a thermodynamically nonfavoured process. Moreover by using a truncated adrenodoxin form with decreased reduction potential as compared with the wild-type ferredoxin-NADP reductase is reduced. Finally these reactions have also been studied using several ferredoxin-NADP reductase mutants at positions crucial for interaction with its physiological partner ferredoxin. The effects observed in their reactions with adrenodoxin do not correlate with those reported for their reactions with ferredoxin. In summary our data indicate that although electron transfer can be achieved

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