Single channel study of the spasmodic mutation α1A52S in recombinant rat glycine receptors

Inherited defects in glycine receptors lead to hyperekplexia, or startle disease. A mutant mouse, spasmodic, that has a startle phenotype, has a point mutation (A52S) in the glycine receptor α1 subunit. This mutation reduces the sensitivity of the receptor to glycine, but the mechanism by which this occurs is not known. We investigated the properties of A52S recombinant receptors by cell-attached patch clamp recording of single-channel currents elicited by 30 – 10000 μM glycine. We used heteromeric receptors, which resemble those found at adult inhibitory synapses. Activation mechanisms were fitted directly to single channel data using the HJCFIT method, which includes an exact correction for missed events. In common with wildtype receptors, only mechanisms with three binding sites and extra shut states could describe the observations. The most physically plausible of these, the ‘flip’ mechanism, suggests that pre-opening isomerisation to the flipped conformation that follows binding is less favoured in mutant than in wild-type receptors, and, especially, that the flipped conformation has a 100-fold lower affinity for glycine than in wildtype receptors. In contrast, the efficacy of the gating reaction was similar to that of wild-type heteromeric receptors. The reduction in affinity for the flipped conformation accounts for the reduction in apparent cooperativity seen in the mutant receptor (without having to postulate interaction between the binding sites) and it accounts for the increased EC50 for responses to glycine that is seen in mutant receptors. This mechanism also predicts accurately the faster decay of synaptic currents that is observed in spasmodic mice

Direct iminization of PEEK

Semi-crystalline poly(ether ketone)s are important high-temperature engineering thermoplastics, but are difficult to characterize at the molecular level because of their insolubility in conventional organic solvents. Here we report that polymers of this type, including PEEK, react cleanly at high temperatures with low-volatility aralkyl amines to afford stable, noncrystalline poly(ether-imine)s, which are readily soluble in solvents such as chloroform, THF and DMF and so characterizable by conventional size-exclusion chromatography

Dibromido[(tert-butyl­amino)dimeth­yl(piperidin-1-ylmeth­yl)silane-κ2 N,N′]zinc(II)

The title compound, [ZnBr2(C12H28N2Si)], is an example of a neutral coordination compound of a bidentate ligand to a metal centre with the Zn atom being coordinated by two Br and two N atoms, yielding a slightly distorted tetra­hedral coordination environment

Υ and Υ′ leptonic widths, abμ, and mb from full lattice QCD

We determine the decay rate to leptons of the ground-state ϒ meson and its first radial excitation in lattice QCD for the first time. We use radiatively improved nonrelativistic QCD for the b quarks and include u, d, s and c quarks in the sea with u=d masses down to their physical values. We find Γðϒ → eþe−Þ ¼ 1.19ð11Þ keV and Γðϒ0 → eþe−Þ ¼ 0.69ð9Þ keV, both in good agreement with experimental results. The decay constants we obtain are included in a summary plot of meson decay constants from lattice QCD given in the Conclusions. We also test time moments of the vector current-current correlator against values determined from the b-quark contribution to σðeþe− → hadronsÞ and calculate the b-quark piece of the hadronic vacuum polarization contribution to the anomalous magnetic moment of the muon, ab μ ¼ 0.271ð37Þ × 10−10. Finally we determine the b-quark mass, obtaining in the MS scheme, ¯ m¯ bðm¯ b; nf ¼ 5Þ ¼ 4.196ð23Þ GeV, the most accurate result from lattice QCD to date

Synthesis, x-ray structure and anion binding properties of a cryptand-like hybrid calixpyrrole

The novel cryptand in/out-3, containing two tripyrrolemethane units briged by three 1,3- diisopropylidenbenzene arms was readily synthesized by a convergent three-step synthesis. It binds fluoride by inclusion with excellent selectivity with respect to a number of other tested anions. The structure of the free receptor and that of its fluoride complex were investigated in solution by NMR spectroscopy. The solid state X-ray structure of the free cryptand 3 was also determined

B-meson decay constants: a more complete picture from full lattice QCD

We extend the picture of $B$-meson decay constants obtained in lattice QCD beyond those of the $B$, $B_s$ and $B_c$ to give the first full lattice QCD results for the $B^*$, $B^*_s$ and $B^*_c$. We use improved NonRelativistic QCD for the valence $b$ quark and the Highly Improved Staggered Quark (HISQ) action for the lighter quarks on gluon field configurations that include the effect of $u/d$, $s$ and $c$ quarks in the sea with $u/d$ quark masses going down to physical values. For the ratio of vector to pseudoscalar decay constants, we find $f_{B^*}/f_B$ = 0.941(26), $f_{B^*_s}/f_{B_s}$ = 0.953(23) (both $2\sigma$ less than 1.0) and $f_{B^*_c}/f_{B_c}$ = 0.988(27). Taking correlated uncertainties into account we see clear indications that the ratio increases as the mass of the lighter quark increases. We compare our results to those using the HISQ formalism for all quarks and find good agreement both on decay constant values when the heaviest quark is a $b$ and on the dependence on the mass of the heaviest quark in the region of the $b$. Finally, we give an overview plot of decay constants for gold-plated mesons, the most complete picture of these hadronic parameters to date.Comment: 20 pages, 9 figures. Minor updates to the discussion in several places and some additional reference

Conformational modulation of sequence recognition in synthetic macromolecules

The different triplet sequences in high molecular weight aromatic copolyimides comprising pyromellitimide units ("I") flanked by either ether-ketone ("K") or ether-sulfone residues ("S") show different binding strengths for pyrene-based tweezer-molecules. Such molecules bind primarily to the diimide unit through complementary π-π-stacking and hydrogen bonding. However, as shown by the magnitudes of 1H NMR complexation shifts and tweezer-polymer binding constants, the triplet "SIS" binds tweezer-molecules more strongly than "KIS" which in turn bind such molecules more strongly than "KIK". Computational models for tweezer-polymer binding, together with single-crystal X-ray analyses of tweezer-complexes with macrocyclic ether-imides, reveal that the variations in binding strength between the different triplet sequences arise from the different conformational preferences of aromatic rings at diarylketone and diarylsulfone linkages. These preferences determine whether or not chain-folding and secondary π−π-stacking occurs between the arms of the tweezermolecule and the 4,4'-biphenylene units which flank the central diimide residue

The response of the tandem pore potassium channel TASK-3 (K2P9.1) to voltage : gating at the cytoplasmic mouth

Although the tandem pore potassium channel TASK-3 is thought to open and shut at its selectivity filter in response to changes of extracellular pH, it is currently unknown whether the channel also shows gating at its inner, cytoplasmic mouth through movements of membrane helices M2 and M4.We used two electrode voltage clamp and single channel recording to show that TASK-3 responds to voltage in a way that reveals such gating. In wild-type channels, Popen was very low at negative voltages, but increased with depolarisation. The effect of voltage was relatively weak and the gating charge small, ∼0.17.Mutants A237T (in M4) and N133A (in M2) increased Popen at a given voltage, increasing mean open time and the number of openings per burst. In addition, the relationship between Popen andvoltagewas shifted to lesspositive voltages. Mutation of putative hinge glycines (G117A, G231A), residues that are conserved throughout the tandem pore channel family, reduced Popen at a given voltage, shifting the relationship with voltage to a more positive potential range. None of these mutants substantially affected the response of the channel to extracellular acidification. We have used the results from single channel recording to develop a simple kinetic model to show how gating occurs through two classes of conformation change, with two routes out of the open state, as expected if gating occurs both at the selectivity filter and at its cytoplasmic mouth