Variable coordination geometries at the diiron(II) active site of ribonucleotide reductase R2

The R2 subunit of Escherichia coli ribonucleotide reductase contains a dinuclear iron center that generates a catalytically essential stable tyrosyl radical by one electron oxidation of a nearby tyrosine residue. After acquisition of Fe(II) ions by the apo protein, the resulting diiron(II) center reacts with O(2) to initiate formation of the radical. Knowledge of the structure of the reactant diiron(II) form of R2 is a prerequisite for a detailed understanding of the O(2) activation mechanism. Whereas kinetic and spectroscopic studies of the reaction have generally been conducted at pH 7.6 with reactant produced by the addition of Fe(II) ions to the apo protein, the available crystal structures of diferrous R2 have been obtained by chemical or photoreduction of the oxidized diiron(III) protein at pH 5-6. To address this discrepancy, we have generated the diiron(II) states of wildtype R2 (R2-wt), R2-D84E, and R2-D84E/W48F by infusion of Fe(II) ions into crystals of the apo proteins at neutral pH. The structures of diferrous R2-wt and R2-D48E determined from these crystals reveal diiron(II) centers with active site geometries that differ significantly from those observed in either chemically or photoreduced crystals. Structures of R2-wt and R2-D48E/W48F determined at both neutral and low pH are very similar, suggesting that the differences are not due solely to pH effects. The structures of these "ferrous soaked" forms are more consistent with circular dichroism (CD) and magnetic circular dichroism (MCD) spectroscopic data and provide alternate starting points for consideration of possible O(2) activation mechanisms.     

Stöber synthesis of monodispersed luminescent silica nanoparticles for bioanalytical assays

We have developed a simple method to prepare bright and photostable luminescent silica nanoparticles of different sizes and narrow size distribution in high yield. The method is based on the use of St?ber synthesis in the presence of a fluorophore to form bright silica nanoparticles. Unlike micro-emulsion-based methods often used to prepare luminescent silica particles, the St?ber method is a one-pot synthesis that is carried out at room temperature under alkaline conditions in ethanol:water mixtures and avoids the use of potentially toxic organic solvents and surfactants. Our luminescent particles contained the transition metal complex tris(1,10-phenanthroline) ruthenium(II) chloride, [Ru(phen)3]Cl2. They showed higher photostability and a longer fluorescence lifetime compared to free Ru(phen)3 solutions. Leakage of dye molecules from the silica particles was negligible, which was attributed to strong electrostatic attractions between the positively charged ruthenium complex and the negatively charged silica. To demonstrate the utility of the highly luminescent silica nanoparticles in bioassays, we further modified their surface with streptavidin and demonstrated their binding to biotinylated glass slides. The study showed that digital counting of the luminescent nanoparticles could be used as an attractive alternative to detection techniques involving analogue luminescence detection in bioanalytical assays.     

Nanoscale fluorescent sensors for intracellular analysis

There is a growing interest in the development of submicron optochemical sensing devices. Miniaturization of sensors to nano-dimensions decreases their typical response time down to the millisecond time scale. Their penetration volume is reduced to a few cubic micrometers and they exhibit a spatial resolution at the nanometer scale. In this review the fabrication of submicron optical fiber fluorescent sensors and particle-based fluorescent nanosensors is described. The functional characteristics of these exciting miniaturized fluorescent sensors and their applications for quantitative measurement of intracellular analytes are demonstrated.     

Regge couplings and intercepts from the planar dual bootstrap

Planar unitarization of dual models is examined in terms of renormalized diagrams. The shift of the Reggeon intercept 1 ? α(0) and the renormalized coupling $\text{g}{}^2\text{N}\text{16π}$ are both expected of order unity in a model with SU(N) Chan-Paton factors. In four space-time dimensions Regge behaviour makes the shift of the dual photon mass reminiscent of Schwinger's mechanism.     

Effects of ion motion in intense beam-driven plasma wakefield accelerators

Recent proposals for using plasma wakefield accelerators (PWFA) as a component of a linear collider have included intense electron beams with densities many times in excess of the plasma density. The beam's electric fields expel the plasma electrons from the beam path to many beam radii in this regime. We analyze here the motion of plasma ions under the beam fields, and find for a proposed PWFA collider scenario that the ions completely collapse inside of the beam. Simulations of ion collapse are presented. Implications of ion motion on the feasibility of the PWFA-based colliders are discussed.