Cfr and RlmN contain a single [4Fe-4S] cluster, which directs two distinct reactivities for S-adenosylmethionine: methyl transfer by SN2 displacement and radical generation

The radical SAM (RS) proteins RlmN and Cfr catalyze methylation of carbons 2 and 8, respectively, of adenosine 2503 in 23S rRNA. Both reactions are similar in scope, entailing the synthesis of a methyl group partially derived from S-adenosylmethionine (SAM) onto electrophilic sp(2)-hybridized carbon atoms via the intermediacy of a protein S-methylcysteinyl (mCys) residue. Both proteins contain five conserved Cys residues, each required for turnover. Three cysteines lie in a canonical RS CxxxCxxC motif and coordinate a [4Fe-4S]-cluster cofactor; the remaining two are at opposite ends of the polypeptide. Here we show that each protein contains only the one "radical SAM" [4Fe-4S] cluster and the two remaining conserved cysteines do not coordinate additional iron-containing species. In addition, we show that, while wild-type RlmN bears the C355 mCys residue in its as-isolated state, RlmN that is either engineered to lack the [4Fe-4S] cluster by substitution of the coordinating cysteines or isolated from Escherichia coli cultured under iron-limiting conditions does not bear a C355 mCys residue. Reconstitution of the [4Fe-4S] cluster on wild-type apo RlmN followed by addition of SAM results in rapid production of S-adenosylhomocysteine (SAH) and the mCys residue, while treatment of apo RlmN with SAM affords no observable reaction. These results indicate that in Cfr and RlmN, SAM bound to the unique iron of the [4Fe-4S] cluster displays two reactivities. It serves to methylate C355 of RlmN (C338 of Cfr), or to generate the 5'-deoxyadenosyl 5'-radical, required for substrate-dependent methyl synthase activity.     

Reactions of HCl and D2O with molten alkali carbonates

The acidic oxide SO? and protic acid HCl are among the gases released in the combustion of coal and the incineration of municipal waste. They are typically removed by wet or dry scrubbing involving calcium carbonate or calcium hydroxide. The molten alkali carbonate eutectic provides a liquid-state alternative that readily absorbs SO? and HCl and does not become covered with a passivating layer. Gas-liquid scattering experiments utilizing the eutectic mixture (44 mol % Li?CO?, 31 mol % Na?CO?, 25 mol % K?CO?) reveal that the reaction probability for HCl(g) + CO?2? → CO?(g) + OH? + Cl? is 0.31 ± 0.02 at 683 K and rises to 0.39 at 783 K. Gaseous CO? is formed within 10?? s or less, implying that the reaction takes place in a liquid depth of less than 1000 ?. When the melt is exposed to D?O, the analogous reaction D?O(g) + CO?2? → CO?(g) + 2OD? occurs too slowly to measure and no water uptake is observed. Together with previous studies of SO?(g) + CO?2? → CO?(g) + SO?2?, these results demonstrate that molten carbonates efficiently remove both gaseous HCl and SO? while reacting at most weakly with water vapor. The experiments further highlight the remarkable ability of hot CO?2? ions to behave as a base in reactions with protic and Lewis acids.     

Minimization of interface roughness in laser deposited Fe/MgO multilayers

The physical properties of multilayers with layer periods in the nanometer range crucially depend on the nature of their interfaces. In this contribution we show how the interfacial roughness of multilayers consisting of two materials with a high difference in specific surface free energy (here MgO/Fe) and thus different growth modes (Volmer–Weber island growth for Fe and layer-by-layer growth for MgO) can be minimized. For this, the layer thickness of the island former has to be large enough that almost all island zipping processes have already occurred. At this thickness the layer is just closed and the intrinsic roughness is at a minimal value. To determine this optimal thickness, in-situ stress measurements as performed in this work are one method of choice. The layer-by-layer growth of the other material with lower surface free energy additionally leads to a further smoothening of the surface. Due to this alternating roughening and smoothing processes, cumulative roughness can be avoided and overall a minimal roughness of the multilayer structure is obtained.     

Development of laser deposited multilayer zone plate structures for soft X-ray radiation

As a novel approach, the combination of pulsed laser deposition and focused ion beam was applied to fabricate different types of multilayer zone plate structures for soft X-ray applications. For this purpose, high quality non-periodic ZrO₂/Ti multilayers were deposited by pulsed laser deposition on planar Si substrates and on rotating steel wires with layer thicknesses according to the Fresnel zone plate law. Linear focusing optics were fabricated by cutting slices out of the multilayers by focused ion beam and placing them directly over pinholes within Si₃N₄ substrates. Additionally, it was shown that laser deposition of depth-graded multilayers on a wire is also a promising way for building up multilayer zone plates with point focus. First experiments using a table-top X-ray source based on a laser-induced plasma show that the determined focal length and spatial resolution of the fabricated multilayer Laue lens corresponds to the designed optic.     

A microfluidic method to study demulsification kinetics

We present the results of experiments studying droplet coalescence in a dense layer of emulsion droplets using microfluidic circuits. The microfluidic structure allows direct observation of collisions and coalescence events between oil droplets dispersed in water. The coalescence rate of a flowing hexadecane-in-water emulsion was measured as a function of the droplet velocity and droplet concentration from image sequences measured with a high-speed camera. A trajectory analysis of colliding droplet pairs allows evaluation of the film drainage profile and coalescence time t(c.) The coalescence times obtained for thousands of droplet pairs enable us to calculate coalescence time distributions for each set of experimental parameters, which are the mean droplet approach velocity (v(0)), the mean dispersed phase fraction (φ) and the mean hydraulic diameter of a droplet pair (d(p)). The expected value E(t(c)) of the coalescence time distributions scales as E(t(c)) is proportional to (v(0))(-0.105±0.043)(d(p))(0.562±0.287), but is independent of φ. We discuss the potential of the procedure for the prediction of emulsion stability in industrial applications.     

Chemical potential of a nonionic surfactant in solution

The chemical potential of a surfactant in solution can be calculated from the Gibbs adsorption equation when the surface excess of the surfactant and the surface tension of the solution as a function of surfactant concentration are known. We have investigated a solution of the nonionic surfactant 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) in the polar solvent 3-hydroxypropionitrile at concentrations below and above the critical micelle concentration (cmc). Neutral impact collision ion scattering spectroscopy was applied for the direct measurement of the surface excess of POPC as a function of concentration. The Gibbs adsorption equation was applied in conjunction with surface tension measurements to evaluate the chemical potential and the activity coefficients of POPC, respectively. We find that the solution shows ideal behavior up to the cmc and that the chemical potential remains constant at concentrations larger than the cmc.