Radical formation in cytochrome c oxidase

AbstractThe formation of radicals in bovine cytochrome c oxidase (bCcO), during the O2 redox chemistry and proton translocation, is an unresolved controversial issue. To determine if radicals are formed in the catalytic reaction of bCcO under single turnover conditions, the reaction of O2 with the enzyme, reduced by either ascorbate or dithionite, was initiated in a custom-built rapid freeze quenching (RFQ) device and the products were trapped at 77K at reaction times ranging from 50μs to 6ms. Additional samples were hand mixed to attain multiple turnover conditions and quenched with a reaction time of minutes. X-band (9GHz) continuous wave electron paramagnetic resonance (CW-EPR) spectra of the reaction products revealed the formation of a narrow radical with both reductants. D-band (130GHz) pulsed EPR spectra allowed for the determination of the g-tensor principal values and revealed that when ascorbate was used as the reductant the dominant radical species was localized on the ascorbyl moiety, and when dithionite was used as the reductant the radical was the SO2− ion. When the contributions from the reductants are subtracted from the spectra, no evidence for a protein-based radical could be found in the reaction of O2 with reduced bCcO. As a surrogate for radicals formed on reaction intermediates, the reaction of hydrogen peroxide (H2O2) with oxidized bCcO was studied at pH 6 and pH 8 by trapping the products at 50μs with the RFQ device to determine the initial reaction events. For comparison, radicals formed after several minutes of incubation were also examined, and X-band and D-band analysis led to the identification of radicals on Tyr-244 and Tyr-129. In the RFQ measurements, a peroxyl (ROO) species was formed, presumably by the reaction between O2 and an amino acid-based radical. It is postulated that Tyr-129 may play a central role as a proton loading site during proton translocation by ejecting a proton upon formation of the radical species and then becoming reprotonated during its reduction via a chain of three water molecules originating from the region of the propionate groups of heme a3. This article is part of a Special Issue entitled: “Allosteric cooperativity in respiratory proteins”

Core binding factors are necessary for natural killer cell development, and cooperate with Notch signaling during T cell specification

CBF{beta} is the non-DNA binding subunit of the core binding factors (CBFs). Mice with reduced CBF{beta} levels display profound, early defects in T but not B cell development. Here we show that CBF{beta} is also required at very early stages of natural killer (NK) cell development. We also demonstrate that T cell development aborts during specification, as the expression of Gata3 and Tcf7, which encode key regulators of T lineage specification, is substantially reduced, as are functional thymic progenitors. Constitutively active Notch or IL-7 signaling cannot restore T cell expansion or differentiation of CBF{beta} insufficient cells, nor can overexpression of Runx1 or CBF{beta} overcome a lack of Notch signaling. Therefore the ability of the prethymic cell to respond appropriately to Notch is dependent on CBF{beta}, and both signals converge to activate the T cell developmental program

Optical Investigations on In0.11Ga0.89N based LEDs Grown on Si (111) Substrate with Different Superlattices Stack Layer Structure

Growing lnxGa1-xN based LEOs on Si is considered challenging as the large lattice mismatch between nitrides and Si material would lead to cracks and defects in the nitride layers. This reduces the luminescence efficiency of the devices. Therefore, in this work, we investigate the effect of the inserting different intermediate structure as an effort to reduce the defects from propagating into the multi-quantum wells (MQWs). Here, In0.11Ga0.89N based LEOs grown on Si (111) substrate with AIN/GaN SLS. In between the LEOs and the SLS, intermediate layers were grown in different structure and in different devices. The idea is to further minimize the impact of the defects propagation of defects and cracks into the MOWs region. We found the lno_,,Gao.asN based LEDs with the insertion of AIGaN/GaN SLS exhibits the best internal quantum efficiency than other devices

Bhlhe40 controls cytokine production by T cells and is essential for pathogenicity in autoimmune neuroinflammation

T(H)1 and T(H)17 cells mediate neuroinflammation in experimental autoimmune encephalomyelitis (EAE), a mouse model of multiple sclerosis. Pathogenic T(H) cells in EAE must produce the pro-inflammatory cytokine granulocyte-macrophage colony stimulating factor (GM-CSF). T(H) cell pathogenicity in EAE is also regulated by cell-intrinsic production of the immunosuppressive cytokine interleukin 10 (IL-10). Here, we demonstrate that mice deficient for the basic helix-loop-helix (bHLH) transcription factor Bhlhe40 (Bhlhe40(−/−)) are resistant to the induction of EAE. Bhlhe40 is required in vivo in a T cell-intrinsic manner, where it positively regulates the production of GM-CSF and negatively regulates the production of IL-10. In vitro, GM-CSF secretion is selectively abrogated in polarized Bhlhe40(−/−) T(H)1 and T(H)17 cells, and these cells show increased production of IL-10. Blockade of IL-10 receptor in Bhlhe40(−/−) mice renders them susceptible to EAE. These findings identify Bhlhe40 as a critical regulator of autoreactive T cell pathogenicity

Discrete approaches to quantum gravity in four dimensions

The construction of a consistent theory of quantum gravity is a problem in theoretical physics that has so far defied all attempts at resolution. One ansatz to try to obtain a non-trivial quantum theory proceeds via a discretization of space-time and the Einstein action. I review here three major areas of research: gauge-theoretic approaches, both in a path-integral and a Hamiltonian formulation, quantum Regge calculus, and the method of dynamical triangulations, confining attention to work that is strictly four-dimensional, strictly discrete, and strictly quantum in nature.Comment: 33 pages, invited contribution to Living Reviews in Relativity; the author welcomes any comments and suggestion

Formation and stability of binary complexes of divalent ecotoxic ions (Ni, Cu, Zn, Cd, Pb) with biodegradable aminopolycarboxylate chelants (dl-2-(2-carboxymethyl)nitrilotriacetic acid, GLDA, and 3-hydroxy-2,2′- iminodisuccinic acid, HIDS) in aqueous solutions

The protonation and complex formation equilibria of two biodegradable aminopolycarboxylate chelants {dl-2-(2-carboxymethyl)nitrilotriacetic acid (GLDA) and 3-hydroxy-2,2′-iminodisuccinic acid (HIDS)} with Ni 2+, Cu2+, Zn2+, Cd2+ and Pb 2+ ions were investigated using the potentiometric method at a constant ionic strength of I = 0.10 mol·dm-3 (KCl) in aqueous solutions at 25 ± 0.1 C. The stability constants of the proton-chelant and metal-chelant species for each metal ion were determined, and the concentration distributions of various complex species in solution were evaluated for each ion. The stability constants (log10 K ML) of the complexes containing Ni2+, Cu2+, Zn2+, Cd2+ and Pb2+ ions followed the identical order of log10 K CuL > log10 K NiL > log10 K PbL > log10 K ZnL > log10 K CdL for either GLDA (13.03 > 12.74 > 11.60 > 11.52 > 10.31) or HIDS (12.63 > 11.30 > 10.21 > 9.76 > 7.58). In each case, the constants obtained for metal-GLDA complexes were larger than the corresponding constants for metal-HIDS complexes. The conditional stability constants (log10 $$K-{\text{ML}}^{\u27}$$) of the metal-chelant complexes containing GLDA and HIDS were calculated in terms of pH, and compared with the stability constants for EDTA and other biodegradable chelants. © 2012 Springer Science+Business Media New York

Comment on "Drug Screening for ALS Using Patient-Specific Induced Pluripotent Stem Cells"

Egawa et al. recently showed the value of patient-specific induced pluripotent stem cells (iPSCs) for modeling amyotrophic lateral sclerosis in vitro. Their study and our work highlight the need for complementary assays to detect small, but potentially important, phenotypic differences between control iPSC lines and those carrying disease mutations

Photoproduction of pions and properties of baryon resonances from a Bonn-Gatchina partial wave analysis

Masses, widths and photocouplings of baryon resonances are determined in a coupled-channel partial wave analysis of a large variety of data. The Bonn-Gatchina partial wave formalism is extended to include a decomposition of t- and u-exchange amplitudes into individual partial waves. The multipole transition amplitudes for $\gamma p\to p\pi^0$ and $\gamma p\to n\pi^+$ are given and compared to results from other analyses.Comment: 18 pages, 14 figure