|Title||A Self-Assembled Respiratory Chain that Catalyzes NADH Oxidation by Ubiquinone-10 Cycling between Complex I and the Alternative Oxidase.|
|Publication Type||Journal Article|
|Year of Publication||2016|
|Authors||J Y Jones, A, Blaza, JN, Bridges, HR, May, B, Moore, AL, Hirst, J|
|Journal||Angew Chem Int Ed Engl|
|Date Published||2016 Jan 11|
Complex I is a crucial respiratory enzyme that conserves the energy from NADH oxidation by ubiquinone-10 (Q10 ) in proton transport across a membrane. Studies of its energy transduction mechanism are hindered by the extreme hydrophobicity of Q10 , and they have so far relied on native membranes with many components or on hydrophilic Q10 analogues that partition into membranes and undergo side reactions. Herein, we present a self-assembled system without these limitations: proteoliposomes containing mammalian complex I, Q10 , and a quinol oxidase (the alternative oxidase, AOX) to recycle Q10 H2 to Q10 . AOX is present in excess, so complex I is completely rate determining and the Q10 pool is kept oxidized under steady-state catalysis. The system was used to measure a fully-defined KM value for Q10 . The strategy is suitable for any enzyme with a hydrophobic quinone/quinol substrate, and could be used to characterize hydrophobic inhibitors with potential applications as pharmaceuticals, pesticides, or fungicides.
|Alternate Journal||Angew. Chem. Int. Ed. Engl.|