Catalytic Asymmetry in Homodimeric H+-Pumping Membrane Pyrophosphatase Demonstrated by Non-Hydrolyzable Pyrophosphate Analogs

Membrane-bound inorganic pyrophosphatase (mPPase) resembles the F-ATPase in catalyzing polyphosphate-energized H+ and Na+ transport across lipid membranes, but differs structurally and mechanistically. Homodimeric mPPase likely uses a “direct coupling” mechanism, in which the proton generated from the water nucleophile at the entrance to the ion conductance channel is transported across the membrane or triggers Na+ transport. The structural aspects of this mechanism, including subunit cooperation, are still poorly understood. Using a refined enzyme assay, we examined the inhibition of K+-dependent H+-transporting mPPase from Desulfitobacterium hafniensee by three non-hydrolyzable PPi analogs (imidodiphosphate and C-substituted bisphosphonates). The kinetic data demonstrated negative cooperativity in inhibitor binding to two active sites, and reduced active site performance when the inhibitor or substrate occupied the other active site. The nonequivalence of active sites in PPi hydrolysis in terms of the Michaelis constant vanished at a low (0.1 mM) concentration of Mg2+ (essential cofactor). The replacement of K+, the second metal cofactor, by Na+ increased the substrate and inhibitor binding cooperativity. The detergent-solubilized form of mPPase exhibited similar active site nonequivalence in PPi hydrolysis. Our findings support the notion that the mPPase mechanism combines Mitchell’s direct coupling with conformational coupling to catalyze cation transport across the membrane.

Coupling a weather model directly to GNSS orbit determination

Neutral atmosphere bends and delays propagation of microwave signals in satellite-based navigation. Weather prediction models can be used to estimate these effects by providing 3-dimensional refraction fields to estimate signal delay in the zenith direction and determine a low-dimensional mapping of this delay to desired azimuth and elevation angles. In this study, a global numerical weather prediction model (OpenIFS licensed for Academic use by ECMWF) is used to generate the refraction fields. The ray-traced slant delays are supplied as such – in contrast to mapping – for an orbit solver (GROOPS software toolkit of TUG) which applies the raw observation method. Here we show that such a close coupling is possible without need for major additional modifications in the solver codes. The main finding here is that the adopted approach provides a very good a priori model for the atmospheric effects on navigation signals, as measured with the midnight discontinuity of GNSS satellite orbits. Our interpretation is that removal of the intermediate mapping step allows to take advantage of the local refraction field asymmetries in the GNSS signal processing. Moreover, the direct coupling helps in identifying deficiencies in the slant delay computation because the modelling errors are not convoluted in the precision-reducing mapping. These conclusions appear robust, despite the relatively small data set of raw code and phase observations covering the core network of 66 ground-based stations of the International GNSS Service over one-month periods in December 2016 and June 2017. More generally, the new configuration enhances our control of geodetic and meteorological aspects of the orbit problem. This is pleasant because we can, for instance, regulate at will the weather model output frequency and increase coverage of spatio-temporal aspects of weather variations. The direct coupling of a weather model in precise GNSS orbit determination presented in this paper provides a unique framework for benefiting even more widely than previously the apparent synergies in space geodesy and meteorology.

Higgs-like spectator field as the origin of structure

We show that the observed primordial perturbations can be entirely sourced by a light spectator scalar field with a quartic potential, akin to the Higgs boson, provided that the field is sufficiently displaced from vacuum during inflation. The framework relies on the indirect modulation of reheating, which is implemented without any direct coupling between the spectator field and the inflaton and does not require non-renormalisable interactions. The scenario gives rise to local non-Gaussianity with $$f_{\mathrm{NL}}\simeq 5$$ f NL ≃ 5 as the typical signal. As an example model where the indirect modulation mechanism is realised for the Higgs boson, we study the Standard Model extended with right-handed neutrinos. For the Standard Model running we find, however, that the scenario analysed does not seem to produce the observed perturbation.