Proton NMR Spectroscopy as a Probe of Dinuclear Copper(II) Active Sites in Metalloproteins. Characterization of the Hyperactive Copper(II)-Substituted Aminopeptidase from \u3cem\u3eAeromonas proteolytica\u3c/em\u3e

Proton NMR spectra of the hyperactive Cu(II)-substituted aminopeptidase from Aeromonas proteolytica (AAP) were recorded in both H2O and D2O buffered solution at pH 6.7. Several remarkably sharp, well resolved hyperfine shifted 1H NMR signals were observed in the 70 to −20 ppm chemical shift range. That hyperfine shifted signals were observed is due to spin-coupling of the two Cu(II) ions. Comparison of the spectra recorded in H2O and D2O buffered solutions indicated that the signals at 44.6, 43.3, and 17.7 ppm were solvent exchangeable. The two most strongly downfield shifted signals were assigned to imidazole N−H protons of the two coordinated histidine residues, while the remaining exchangeable signal was assigned to a peptidyl N−H proton that is in close proximity to the dicopper(II) center. One-dimensional NOE studies at pH 6.7 revealed two Y−CH2−CH\u3c moieties that were assigned to coordinated aspartic acid and histidine residues. In addition, a Y−CH2−CH2−CH\u3c moiety was also identified and was assigned to the coordinated glutamic acid residue, Glu152. All of the hyperfine shifted signals for [CuCu(AAP)] sharpened and shifted toward the diamagnetic region as the temperature was increased following Curie behavior. Fits of these data and those of a series of magnetically diverse μ-phenoxo and μ-alkoxo dicopper(II) model complexes to the population distribution of the ground and first excited states, provided information on the magnetic properties of dicopper(II) clusters. These fits indicated that the two Cu(II) ions in AAP are ferromagnetically coupled with a 2J value of 50 + 40 cm-1. These data provide the first structural information regarding the hyperactive [CuCu(AAP)] enzyme and are discussed in terms of the previously proposed mechanism of action for AAP

Static Quark Potential and the Renormalized Anisotropy on Tadpole Improved Anisotropic Lattices

Static quark potential is studied using a tadpole improved gauge lattice action. The scale is set using the potential for a wide range of bare parameters. The renormalized anisotropy of the lattice is also measured.Comment: 11 pages, 5 figures, accepted for publication in Int. J. Mod. Phys.

Probing Half-odd Topological Number with Cold Atoms in a Non-Abelian Optical Lattice

We propose an experimental scheme to probe the contribution of a single Dirac cone to the Hall conductivity as half-odd topological number sequence. In our scheme, the quantum anomalous Hall effect as in graphene is simulated with cold atoms trapped in an optical lattice and subjected to a laser-induced non-Abelian gauge field. By tuning the laser intensity to change the gauge flux, the energies of the four Dirac points in the first Brillouin zone are shifted with each other and the contribution of the single Dirac cone to the total atomic Hall conductivity is manifested. We also show such manifestation can be experimentally probed with atomic density profile measurements.Comment: 5 pages, 3 figure

A Bayesian Approach to Estimate the Size and Structure of the Broad-line Region in Active Galactic Nuclei Using Reverberation Mapping Data

This is the first paper in a series devoted to systematic study of the size and structure of the broad-line region (BLR) in active galactic nuclei (AGNs) using reverberation mapping (RM) data. We employ a recently developed Bayesian approach that statistically describes the variabibility as a damped random walk process and delineates the BLR structure using a flexible disk geometry that can account for a variety of shapes, including disks, rings, shells, and spheres. We allow for the possibility that the line emission may respond non-linearly to the continuum, and we detrend the light curves when there is clear evidence for secular variation. We use a Markov Chain Monte Carlo implementation based on Bayesian statistics to recover the parameters and uncertainties for the BLR model. The corresponding transfer function is obtained self-consistently. We tentatively constrain the virial factor used to estimate black hole masses; more accurate determinations will have to await velocity-resolved RM data. Application of our method to RM data with Hbeta monitoring for about 40 objects shows that the assumed BLR geometry can reproduce quite well the observed emission-line fluxes from the continuum light curves. We find that the Hbeta BLR sizes obtained from our method are on average ~20% larger than those derived from the traditional cross-correlation method. Nevertheless, we still find a tight BLR size-luminosity relation with a slope of alpha=0.55\pm0.03 and an intrinsic scatter of ~0.18 dex. In particular, we demonstrate that our approach yields appropriate BLR sizes for some objects (such as Mrk 142 and PG 2130+099) where traditional methods previously encountered difficulties.Comment: 17 pages, 10 figures, 2 tables; minor reversion to match the published versio

Cyclin B1/Cdk1 phosphorylation of mitochondrial p53 induces anti-apoptotic response.

The pro-apoptotic function of p53 has been well defined in preventing genomic instability and cell transformation. However, the intriguing fact that p53 contributes to a pro-survival advantage of tumor cells under DNA damage conditions raises a critical question in radiation therapy for the 50% human cancers with intact p53 function. Herein, we reveal an anti-apoptotic role of mitochondrial p53 regulated by the cell cycle complex cyclin B1/Cdk1 in irradiated human colon cancer HCT116 cells with p53(+/+) status. Steady-state levels of p53 and cyclin B1/Cdk1 were identified in the mitochondria of many human and mouse cells, and their mitochondrial influx was significantly enhanced by radiation. The mitochondrial kinase activity of cyclin B1/Cdk1 was found to specifically phosphorylate p53 at Ser-315 residue, leading to enhanced mitochondrial ATP production and reduced mitochondrial apoptosis. The improved mitochondrial function can be blocked by transfection of mutant p53 Ser-315-Ala, or by siRNA knockdown of cyclin B1 and Cdk1 genes. Enforced translocation of cyclin B1 and Cdk1 into mitochondria with a mitochondrial-targeting-peptide increased levels of Ser-315 phosphorylation on mitochondrial p53, improved ATP production and decreased apoptosis by sequestering p53 from binding to Bcl-2 and Bcl-xL. Furthermore, reconstitution of wild-type p53 in p53-deficient HCT116 p53(-/-) cells resulted in an increased mitochondrial ATP production and suppression of apoptosis. Such phenomena were absent in the p53-deficient HCT116 p53(-/-) cells reconstituted with the mutant p53. These results demonstrate a unique anti-apoptotic function of mitochondrial p53 regulated by cyclin B1/Cdk1-mediated Ser-315 phosphorylation in p53-wild-type tumor cells, which may provide insights for improving the efficacy of anti-cancer therapy, especially for tumors that retain p53