Phosphorylation Mechanism of Phosphomevalonate Kinase: Implications for Rational Engineering of Isoprenoid Biosynthetic Pathway Enzymes

The mevalonate pathway is of important clinical, pharmaceutical, and biotechnological relevance. However, lack of the understanding of the phosphorylation mechanism of the kinases in this pathway has limited rationally engineering the kinases in industry. Here the phosphorylation reaction mechanism of a representative kinase in the mevalonate pathway, phosphomevalonate kinase, was studied by using molecular dynamics and hybrid QM/MM methods. We find that a conserved residue (Ser106) is reorientated to anchor ATP via a stable H-bond interaction. In addition, Ser213 located on the α-helix at the catalytic site is repositioned to further approach the substrate, facilitating the proton transfer during the phosphorylation. Furthermore, we elucidate that Lys101 functions to neutralize the negative charge developed at the β-, γ-bridging oxygen atom of ATP during phosphoryl transfer. We demonstrate that the dissociative catalytic reaction occurs via a direct phosphorylation pathway. This is the first study on the phosphorylation mechanism of a mevalonate pathway kinase. The elucidation of the catalytic mechanism not only sheds light on the common catalytic mechanism of the GHMP kinase superfamily but also provides the structural basis for engineering the mevalonate pathway kinases to further exploit their applications in the production of a wide range of fine chemicals such as biofuels or pharmaceuticals

New transitions and feeding of the J\u3csup\u3eπ\u3c/sup\u3e=(8\u3csup\u3e+\u3c/sup\u3e) isomer in \u3csup\u3e186\u3c/sup\u3eRe

The spallation neutron source at the Los Alamos Neutron Science Center Weapons Neutron Research facility was used to populate excited states in 186Re via (n,2nγ) reactions on an enriched 187Re target. Gamma rays were detected with the GErmanium Array for Neutron Induced Excitations spectrometer, a Compton-suppressed array of 18 HPGe detectors. Incident neutron energies were determined by the time-of-flight technique and used to obtain γ-ray excitation functions for the purpose of identifying γ rays by reaction channel. Analysis of the singles γ-ray spectrum gated on the neutron energy range 10≤En≤25MeV resulted in five transitions and one level added to the 186Re level scheme. The additions include the placement of three γ rays at 266.7, 381.2, and 647.7 keV which have been identified as feeding the 2.0×105yr, Jπ=(8+) isomer and yield an improved value of 148.2(5)keV for the isomer energy. These transitions may have astrophysical implications related to the use of the Re-Os cosmochronometer. Abstract © APS

The Electronic Structure and Secondary Pyroelectric Properties of Lithium Tetraborate

We review the pyroelectric properties and electronic structure of Li2B4O7(110) and Li2B4O7(100) surfaces. There is evidence for a pyroelectric current along the [110] direction of stoichiometric Li2B4O7 so that the pyroelectric coefficient is nonzero but roughly 103 smaller than along the [001] direction of spontaneous polarization. Abrupt decreases in the pyroelectric coefficient along the [110] direction can be correlated with anomalies in the elastic stiffness C_33^D contributing to the concept that the pyroelectric coefficient is not simply a vector but has qualities of a tensor, as expected. The time dependent surface photovoltaic charging suggests that surface charging is dependent on crystal orientation and doping, as well as temperature

The Electronic Structure and Secondary Pyroelectric Properties of Lithium Tetraborate

We review the pyroelectric properties and electronic structure of Li2B4O7(110) and Li2B4O7(100) surfaces. There is evidence for a pyroelectric current along the [110] direction of stoichiometric Li2B4O7 so that the pyroelectric coefficient is nonzero but roughly 103 smaller than along the [001] direction of spontaneous polarization. Abrupt decreases in the pyroelectric coefficient along the [110] direction can be correlated with anomalies in the elastic stiffness contributing to the concept that the pyroelectric coefficient is not simply a vector but has qualities of a tensor, as expected. The time dependent surface photovoltaic charging suggests that surface charging is dependent on crystal orientation and doping, as well as temperature

Particle-in-cell Simulations of Ion Dynamics in a Pinched-beam Diode

article-in-cell simulations of a 1.6 MV, 800 kA, and 50 ns pinched-beam diode have been completed with emphasis placed on the quality of the ion beams produced. Simulations show the formation of multiple regions in the electron beam flow characterized by locally high charge and current density (“hot spots”). As ions flow through the electron-space-charge cloud, these hot spots electrostatically attract ions to produce a non-uniform ion current distribution. The length of the cavity extending beyond the anode-to-cathode gap (i.e., behind the cathode tip) influences both the number and amplitude of hot spots. A longer cavity length increases the number of hot spots yet significantly reduces the amplitude producing a smoother, more uniform ion beam than for shorter cavities. The net current and the ion bending angles are also significantly smaller with long cavities

Rare Earth Dopant (Nd, Gd, Dy, and Er) Hybridization in Lithium Tetraborate

The four dopants (Nd, Gd, Dy, and Er) substitutionally occupy the Li+ sites in lithium tetraborate (Li2B4O7: RE) glasses as determined by analysis of the extended X-ray absorption fine structure. The dopants are coordinated by 6-8 oxygen at a distance of 2.3 to 2.5 Å, depending on the rare earth. The inverse relationship between the RE-O coordination distance and rare earth (RE) atomic number is consistent with the expected lanthanide atomic radial contraction with increased atomic number. Through analysis of the X-ray absorption near edge structure, the rare earth dopants adopt the RE3+ valence state. There are indications of strong rare earth 5d hybridization with the trigonal and tetrahedral formations of BO3 and BO4 based on the determination of the rare earth substitutional Li+ site occupancy from the X-ray absorption near edge structure data. The local oxygen disorder around the RE3+ luminescence centers evident in the structural determination of the various glasses, and the hybridization of the RE3+ dopants with the host may contribute to the asymmetry evident in the luminescence emission spectral lines. The luminescence emission spectra are indeed characteristic of the expected f-to-f transitions; however, there is an observed asymmetry in some emission lines