title: From in silico to in spectro kinetics of respiratory complex I.
abstract: An enzyme's activity is the consequence of its structure. The stochastic approach we developed to study the functioning of the respiratory complexes is based upon their 3D structure and their physical and chemical properties. Consequently it should predict their kinetic properties. In this paper we compare the predictions of our stochastic model derived for the complex I with a number of experiments performed with a large range of complex I substrates and products. A good fit was found between the experiments and the prediction of our stochastic approach. We show that, due to the spatial separation of the two half redox reactions (NADH/NAD and Q/QH(2)), the kinetics cannot necessarily obey a simple mechanism (ordered or ping-pong for instance). A plateau in the kinetics is observed at high substrates concentrations, well evidenced in the double reciprocal plots, which is explained by the limiting rate of quinone reduction as compared with the oxidation of NADH at the other end of complex I. Moreover, we show that the set of the seven redox reactions in between the two half redox reactions (NADH/NAD and Q/QH(2)) acts as an electron buffer. An inhibition of complex I activity by quinone is observed at high concentration of this molecule, which cannot be explained by a simple stochastic model based on the known structure. We hypothesize that the distance between the catalytic site close to N2 (iron/sulfur redox center that transfers electrons to quinone) and the membrane forces the quinone/quinol to take several positions in between these sites. We represent these possible positions by an extra site necessarily occupied by the quinone/quinol molecules on their way to the redox site. With this hypothesis, we are able to fit the kinetic experiments over a large range of substrates and products concentrations. The slow rate constants derived for the transition between the two sites could be an indication of a conformational change of the enzyme during the quinone/quinol movement. This article is part of a Special Issue entitled: 17th European Bioenergetics Conference (EBEC 2012).
authors.affiliation: Institute of Biochemistry and Genetics of the Cell, Bordeaux cedex, France.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Ransac
authors.forename: Margit
authors.initials: M
authors.lastname: Heiske
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Mazat
mesh_terms: D000818:Animals; D002417:Cattle; D003198:Computer Simulation; D004579:Electron Transport; D042967:Electron Transport Complex I / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008929:Mitochondria, Heart / Q000201:enzymology*; D024101:Mitochondrial Proteins / Q000737:chemistry* / Q000378:metabolism; D008956:Models, Chemical; D009124:Muscle Proteins / Q000737:chemistry* / Q000378:metabolism; D009243:NAD / Q000737:chemistry / Q000378:metabolism; D009249:NADP / Q000737:chemistry / Q000378:metabolism; D010084:Oxidation-Reduction
chemical_list: D024101:Mitochondrial Proteins; D009124:Muscle Proteins; D009243:NAD; D009249:NADP; D042967:Electron Transport Complex I