Variation in the organization and subunit composition of the mammalian pyruvate dehydrogenase complex E2/E3BP core assembly

The final version of this article is available at the link below.Crucial to glucose homoeostasis in humans, the hPDC (human pyruvate dehydrogenase complex) is a massive molecular machine comprising multiple copies of three distinct enzymes (E1–E3) and an accessory subunit, E3BP (E3-binding protein). Its icosahedral E2/E3BP 60-meric ‘core’ provides the central structural and mechanistic framework ensuring favourable E1 and E3 positioning and enzyme co-operativity. Current core models indicate either a 48E2+12E3BP or a 40E2+20E3BP subunit composition. In the present study, we demonstrate clear differences in subunit content and organization between the recombinant hPDC core (rhPDC; 40E2+20E3BP), generated under defined conditions where E3BP is produced in excess, and its native bovine (48E2+12E3BP) counterpart. The results of the present study provide a rational basis for resolving apparent differences between previous models, both obtained using rhE2/E3BP core assemblies where no account was taken of relative E2 and E3BP expression levels. Mathematical modelling predicts that an ‘average’ 48E2+12E3BP core arrangement allows maximum flexibility in assembly, while providing the appropriate balance of bound E1 and E3 enzymes for optimal catalytic efficiency and regulatory fine-tuning. We also show that the rhE2/E3BP and bovine E2/E3BP cores bind E3s with a 2:1 stoichiometry, and propose that mammalian PDC comprises a heterogeneous population of assemblies incorporating a network of E3 (and possibly E1) cross-bridges above the core surface.This work was partly supported by EPSRC (under grants GR/R99393/01 and EP/C015452/1)

Contingent infrastructure and the dilution of ‘Chineseness’: Reframing roads and rail in Kampala and Addis Ababa

Amid growing interest in China’s role in financing and building infrastructure in Africa, there is still little research on how Chinese-financed infrastructures are negotiated and realised at the city and metropolitan scale. We compare the Light Rail Transit in Addis Ababa, Ethiopia with the expressway linking Kampala to Entebbe airport in Uganda, examining the processes of bargaining behind these transport infrastructures and their emergent effects on urban land use and city-dwellers’ mobility. We find that both projects were designed and implemented through opaque negotiations between African national elites and Chinese agencies, with little or no engagement from city authorities, leading to haphazard outcomes that are poorly integrated with broader planning. Yet we also suggest that despite being enabled and mediated by Chinese agencies, such projects do not embody a Chinese global vision. They instead reflect the entrepreneurial activities of Chinese contractors and the varying ways in which these connect with African national governments’ shifting priorities. Moreover, as they are subsumed into the urban context, these transposed infrastructures have been rapidly repurposed and their ‘Chineseness’ diluted, with one morphing into an infrastructure for the poor and the other into a site of private value extraction. We thus argue that, far from representing a domineering or neo-colonial influence, Chinese-financed infrastructures that land in institutionally complex African city-regions can be rapidly swallowed up into the political-economic landscape, producing contingent benefits and disbenefits that are far removed from the visions of any planners – Chinese or African, past or present

Forward K+ production in subthreshold pA collisions at 1.0 GeV

K+ meson production in pA (A = C, Cu, Au) collisions has been studied using the ANKE spectrometer at an internal target position of the COSY-Juelich accelerator. The complete momentum spectrum of kaons emitted at forward angles, theta < 12 degrees, has been measured for a beam energy of T(p)=1.0 GeV, far below the free NN threshold of 1.58 GeV. The spectrum does not follow a thermal distribution at low kaon momenta and the larger momenta reflect a high degree of collectivity in the target nucleus.Comment: 4 pages, 3 figure

Below the Belt? Territory and Development in China’s International Rise

China’s internationalization has been heralded by some as a new era of South–South cooperation. Yet such framings of development are pitched at an abstract space of the ‘global South’ which conceals more than it reveals. With some theory moving towards ontologies of ‘global development’, we need to capture both the connectedness and the local specificity of increasingly diffuse processes. This article sets out a more fine-grained understanding of how political territories and processes are imagined and produced by and through China’s internationalization, focusing on infrastructure as a ‘technology’ of territorialization. Much of the focus on China’s internationalization has been on state-to-state relations, but this obscures the ‘omni-channel politics’ that China practises. Using a critical literature review and illustrative case study, this article develops the idea of omni-channel politics to posit a view of ‘twisted’ territories in which political processes and development outcomes are more complex and contingent

Neurochemical Changes in the Mouse Hippocampus Underlying the Antidepressant Effect of Genetic Deletion of P2X7 Receptors.

Recent investigations have revealed that the genetic deletion of P2X7 receptors (P2rx7) results in an antidepressant phenotype in mice. However, the link between the deficiency of P2rx7 and changes in behavior has not yet been explored. In the present study, we studied the effect of genetic deletion of P2rx7 on neurochemical changes in the hippocampus that might underlie the antidepressant phenotype. P2X7 receptor deficient mice (P2rx7-/-) displayed decreased immobility in the tail suspension test (TST) and an attenuated anhedonia response in the sucrose preference test (SPT) following bacterial endotoxin (LPS) challenge. The attenuated anhedonia was reproduced through systemic treatments with P2rx7 antagonists. The activation of P2rx7 resulted in the concentration-dependent release of [3H]glutamate in P2rx7+/+ but not P2rx7-/- mice, and the NR2B subunit mRNA and protein was upregulated in the hippocampus of P2rx7-/- mice. The brain-derived neurotrophic factor (BDNF) expression was higher in saline but not LPS-treated P2rx7-/- mice; the P2rx7 antagonist Brilliant blue G elevated and the P2rx7 agonist benzoylbenzoyl ATP (BzATP) reduced BDNF level. This effect was dependent on the activation of NMDA and non-NMDA receptors but not on Group I metabotropic glutamate receptors (mGluR1,5). An increased 5-bromo-2-deoxyuridine (BrdU) incorporation was also observed in the dentate gyrus derived from P2rx7-/- mice. Basal level of 5-HT was increased, whereas the 5HIAA/5-HT ratio was lower in the hippocampus of P2rx7-/- mice, which accompanied the increased uptake of [3H]5-HT and an elevated number of [3H]citalopram binding sites. The LPS-induced elevation of 5-HT level was absent in P2rx7-/- mice. In conclusion there are several potential mechanisms for the antidepressant phenotype of P2rx7-/- mice, such as the absence of P2rx7-mediated glutamate release, elevated basal BDNF production, enhanced neurogenesis and increased 5-HT bioavailability in the hippocampus

S. aureus MscL Is a Pentamer In Vivo but of Variable Stoichiometries In Vitro: Implications for Detergent-Solubilized Membrane Proteins

Detergent-induced rearrangements of membrane-protein subunits explain why two MscL channel stoichiometries have been resolved by X-ray crystallography - but S. aureus MscL is truly a pentamer in vivo

Identifying core features of adaptive metabolic mechanisms for chronic heat stress attenuation contributing to systems robustness

The contribution of metabolism to heat stress may play a significant role in defining robustness and recovery of systems; either by providing the energy and metabolites required for cellular homeostasis, or through the generation of protective osmolytes. However, the mechanisms by which heat stress attenuation could be adapted through metabolic processes as a stabilizing strategy against thermal stress are still largely unclear. We address this issue through metabolomic and transcriptomic profiles for populations along a thermal cline where two seagrass species, Zostera marina and Zostera noltii, were found in close proximity. Significant changes captured by these profile comparisons could be detected, with a larger response magnitude observed in northern populations to heat stress. Sucrose, fructose, and myo-inositol were identified to be the most responsive of the 29 analyzed organic metabolites. Many key enzymes in the Calvin cycle, glycolysis and pentose phosphate pathways also showed significant differential expression. The reported comparison suggests that adaptive mechanisms are involved through metabolic pathways to dampen the impacts of heat stress, and interactions between the metabolome and proteome should be further investigated in systems biology to understand robust design features against abiotic stress

Selective disruption of the DNA polymerase III α–β complex by the umuD gene products

DNA polymerase III (DNA pol III) efficiently replicates the Escherichia coli genome, but it cannot bypass DNA damage. Instead, translesion synthesis (TLS) DNA polymerases are employed to replicate past damaged DNA; however, the exchange of replicative for TLS polymerases is not understood. The umuD gene products, which are up-regulated during the SOS response, were previously shown to bind to the α, β and ε subunits of DNA pol III. Full-length UmuD inhibits DNA replication and prevents mutagenic TLS, while the cleaved form UmuD′ facilitates mutagenesis. We show that α possesses two UmuD binding sites: at the N-terminus (residues 1–280) and the C-terminus (residues 956–975). The C-terminal site favors UmuD over UmuD′. We also find that UmuD, but not UmuD′, disrupts the α–β complex. We propose that the interaction between α and UmuD contributes to the transition between replicative and TLS polymerases by removing α from the β clamp

Optimization of radial diffusion coefficients for the proton radiation belt during the CRRES era

Proton flux measurements from the Proton Telescope instrument aboard the CRRES satellite are revisited, and used to drive a radial diffusion model of the inner proton belt at 1.1 ≤ L ≤ 1.65. Our model utilises a physics‐based evaluation of the cosmic ray albedo neutron decay (CRAND) source, and coulomb collisional loss is driven by a drift averaged density model combining results from the International Reference Ionosphere, NRLMSIS‐00 atmosphere and Radio Plasma Imager plasmasphere models, parameterised by solar activity and season. We drive our model using time‐averaged data at L = 1.65 to calculate steady state profiles of equatorial phase space density, and optimise our choice of radial diffusion coefficients based on four defining parameters to minimise the difference between model and data. This is first performed for a quiet period when the belt can be assumed to represent steady state. Additionally, we investigate fitting steady state solutions to time averages taken during active periods where the data exhibits limited deviation from steady state, demonstrated by CRRES measurements following the 24th March 1991 storm. We also discuss a way to make the optimisation process more reliable by excluding periods of variability in plasmaspheric density from any time average. Lastly, we compare our resultant diffusion coefficients to those derived via a similar process in previous work, and diffusion coefficients derived for electrons from ground and in situ observations. We find that higher diffusion coefficients are derived compared with previous work, and suggest more work is required to derive proton diffusion coefficients for different geomagnetic activity levels

China’s Dam Builders: their role in transboundary river management in Southeast Asia

This article investigates China’s role as the world’s largest builder of and investor in large dams, focussing on the Greater Mekong Sub-Region in South-East Asia. It addresses the role Chinese actors play in dam-building as well as the environmental, social, economic and political implications by drawing on case studies from Cambodia and Vietnam. The article finds that China’s dam-building is perceived very differently in different countries of South-East Asia. In Cambodia, the dams in the Greater Mekong Sub-Region are considered instruments of economic growth and development, whereas downstream in Vietnam the dams are seen as potentially undermining national growth, development and security