The proton pumping stoichiometry of purified mitochondrial complex I reconstituted into proteoliposomes

NADH:ubiquinone oxidoreductase (complex I) is the largest and most complicated enzyme of aerobic electron transfer. The mechanism how it uses redox energy to pump protons across the bioenergetic membrane is still not understood. Here we determined the pumping stoichiometry of mitochondrial complex I from the strictly aerobic yeast Yarrowia lipolytica. With intact mitochondria, the measured value of 3.8H(->+)/2e(-) indicated that four protons are pumped per NADH oxidized. For purified complex I reconstituted into proteoliposomes we measured a very similar pumping stoichiometry of 3.6H(->+)/2e(-). This is the first demonstration that the proton pump of complex I stayed fully functional after purification of the enzyme. (c) 2006 Elsevier B.V. All rights reserved.

Proton pumping by complex I (NADH : ubiquinone oxidoreductase) from Yarrowia lipolytica reconstituted into proteoliposomes

The mechanism of energy converting NADH:ubiquinone oxidoreductase (complex 1) is Still unknown. A current controversy centers around the question whether electron transport of complex I is always linked to vectorial proton translocation or whether in some organisms the enzyme pumps sodium ions instead. To develop better experimental tools to elucidate its mechanism, we have reconstituted the affinity purified enzyme into proteoliposomes and monitored the generation of Delta pH and Delta psi. We tested several detergents to solubilize the asolectin used for liposome formation. Tightly coupled proteoliposomes containing highly active complex I were obtained by detergent removal with BioBeads after total solubilization or the phospholipids with n-octyl-beta-D-glucopyranoside. We have used dyes to monitor the formation of the two components of the proton motive force, Delta pH and Delta psi, across the liposomal membrane, and analyzed the effects of inhibitors, uncouplers and ionophores on this process. We show that electron transfer of complex I of the lower eukaryote Y. lipolytica is clearly linked to proton translocation. While this study was not specifically designed to demonstrate possible additional sodium translocating properties of complex 1, we did not find indications for primary or secondary Na+ translocation by Y lipolytica complex I. (c) 2005 Elsevier B.V. All rights reserved.

Laser-driven quasimonoenergetic proton burst from water spray target

A narrow band proton bursts at energies of 1.6 +/- 0.08 MeV were observed when a water spray consisting of empty set(150 nm)-diameter droplets was irradiated by an ultrashort laser pulse of about 45 fs duration and at an intensity of 5 X 10(19) W/cm(2). The results are explained by a Coulomb explosion of sub-laser-wavelength droplets composed of two ion species. The laser prepulse plays an important role. By pre-evaporation of the droplets, its diameter is reduced so that the main pulse can interact with a smaller droplet, and this remaining bulk can be ionized to high states. In the case of water, the mixture of quite differently charged ions establishes an

Tomography of an ultrafast laser driven proton source

Using a multichannel Thomson spectrometer we have implemented a tomographic approach allowing the reconstruction of the emission characteristic of a laser driven proton source with high energy and spatial resolution. The results demonstrate the complexity of the temporal and spatial characteristics of such a source. The emitted proton beam, which is laminar and divergent at high energies, becomes convergent at low energies. This implies that a fraction of the proton beam having this kinetic energy is emitted in a collimated way from the target at the

Proton radiography of a shock-compressed target

In this paper we report on the radiography of a shock-compressed target using laser produced proton beams. A low-density carbon foam target was shock compressed by long pulse high-energy laser beams. The shock front was transversally probed with a proton beam produced in the interaction of a high intensity laser beam with a gold foil. We show that from radiography data, the density profile in the shocked target can be deduced using Monte Carlo simulations. By changing the delay between long and short pulse beams, we could probe different plasma conditions and structures, demonstrating that the details of the steep density gradient can be resolved. This technique is validated as a diagnostic for the investigation of warm dense plasmas, allowing an in situ characterization of high-density contrasted plasmas.

Progress in proton radiography for diagnosis of ICF-relevant plasmas

Proton radiography using laser-driven sources has been developed as a diagnostic since the beginning of the decade, and applied successfully to a range of experimental situations. Multi-MeV protons driven from thin foils via the Target Normal Sheath Acceleration mechanism, offer, under optimal conditions, the possibility of probing laser-plasma interactions, and detecting electric and magnetic fields as well as plasma density gradients with similar to ps temporal resolution and similar to 5-10 mu m spatial resolution. In view of these advantages, the use of proton radiography as a diagnostic in experiments of relevance to Inertial Confinement Fusion is currently considered in the main fusion laboratories. This paper will discuss recent advances in the application of laser-driven radiography to experiments of relevance to Inertial Confinement Fusion. In particular we will discuss radiography of hohlraum and gasbag targets following the interaction of intense ns pulses. These experiments were carried out at the HELEN laser facility at AWE (UK), and proved the suitability of this diagnostic for studying, with unprecedented detail, laser-plasma interaction mechanisms of high relevance to Inertial Confinement Fusion. Non-linear solitary structures of relevance to space physics, namely phase space electron holes, have also been highlighted by the measurements. These measurements are discussed and compared to existing models.

Simulation of a collisionless planar electrostatic shock in a proton-electron plasma with a strong initial thermal pressure change

The localized deposition of the energy of a laser pulse, as it ablates a solid target, introduces high thermal pressure gradients in the plasma. The thermal expansion of this laser-heated plasma into the ambient medium (ionized residual gas) triggers the formation of non-linear structures in the collisionless plasma. Here an electron-proton plasma is modelled with a particle-in-cell simulation to reproduce aspects of this plasma expansion. A jump is introduced in the thermal pressure of the plasma, across which the otherwise spatially uniform temperature and density change by a factor of 100. The electrons from the hot plasma expand into the cold one and the charge imbalance drags a beam of cold electrons into the hot plasma. This double layer reduces the electron temperature gradient. The presence of the low-pressure plasma modifies the proton dynamics compared with the plasma expansion into a vacuum. The jump in the thermal pressure develops into a primary shock. The fast protons, which move from the hot into the cold plasma in the form of a beam, give rise to the formation of phase space holes in the electron and proton distributions. The proton phase space holes develop into a secondary shock that thermalizes the beam.

Quantitative analysis of proton imaging measurements of laser-induced plasmas

A method for obtaining quantitative information about electric field and charge distributions from proton imaging measurements of laser-induced plasmas is presented. A parameterised charge distribution is used as target plasma. The deflection of a proton beam by the electric field of such a plasma is simulated numerically as well as the resulting proton density, which will be obtained on a screen behind the plasma according to the proton imaging technique. The parameters of the specific charge distributions are delivered by a combination of linear regression and nonlinear fitting of the calculated proton density distribution to the measured optical density of a radiochromic film screen changed by proton exposure. It is shown that superpositions of spherical Gaussian charge distributions as target plasma are sufficient to simulate various structures in proton imaging measurements, which makes this method very flexible.

A new algorithm for spectral and spatial reconstruction of proton beams from dosimetric measurements

We report on a new algorithm developed for the dosimetric analysis of broad-spectrum, multi-MeV laser-accelerated proton beams. The algorithm allows the reconstruction of the proton beam spectrum from radiochromic film data. This processing technique makes dosimetry measurements a viable alternative to the use of track detectors for spatially and spectrally resolved proton beam analysis. (C) 2003 Elsevier B.V. All rights reserved.

Proton moire fringes for diagnosing electromagnetic fields in opaque materials and plasmas

High contrast proton moire fringes have been obtained in a laser-produced proton beam. Moire u fringes with modulation of 20%-30% were observed in protons with energies in the range of 4 - 7 MeV. Monte Carlo simulations with simple test fields showed that shifts in the moire u fringes can be used to give quantitative information on the strength of transient electromagnetic fields inside plasmas and materials that are opaque to conventional probing methods. (C) 2003 American Institute of Physics.