Modeling hydrogen exchange of proteins by a multiscale method

We proposed a practical way for mapping the results of coarse-grained molecular simulations to the observables in hydrogen change experiments.By combining an atomic-interaction based coarse-grained model with an all-atom structure reconstruction algorithm,we reproduced the experimental hydrogen exchange data with reasonable accuracy using molecular dynamics simulations.We also showed that the coarse-grained model can be further improved by imposing experimental restraints from hydrogen exchange data via an iterative optimization strategy.These results suggest that it is feasible to develop an integrative molecular simulation scheme by incorporating the hydrogen exchange data into the coarse-grained molecular dynamics simulations and therefore help to overcome the accuracy bottleneck of coarse-grained models.

Resuscitation from Prolonged Ventricular Fibrillation by Epinephrine Combined with Sodium-Hydrogen Exchanger Isoform-1 Inhibitor Cariporide

Objective To test the resuscitative effects from prolonged ventricular fibrillation by epinephrine combined with sodium hydrogen exchanger isoform 1 inhibitor Cariporide. Methods 16 rats were received a 3 mg/kg bolus of Cariporide or the same volume of 0.9%NaCl solution (control) 15 seconds before completion 12 minutes untreated VF. Chest compression (CC) was started for a total of 8 minutes. Adjusted the depth of compressor so that the aortic diastolic pressure to 25～28 mmHg during the 2nd minute of CC. Fix the depth of the piston and this depth was used throughout the remaining 6 minutes of CC. 10 seconds before starting the 3rd minute of chest compression, injected epinephrine (30 μg/kg). Recorded the time at which restoration of spontaneous circulation (ROSC) occurred in Cariporide treated rats. Electrical defibrillation was timed in control group to match the time of spontaneous defibrillation in Cariporide treated rats. To the rats, which cant be defibrillated spontaneously, received chest compression and rescues electrical shocks. Results compared with control group, with the same CC depth, Cariporide treated rats received the higher and longer lasting coronary perfusion pressure (P< 0.05), higher resuscitative rate (P< 0.05), less post resuscitative ventricular ectopic activities (P< 0.001), better hemodynamic effects and longer survival time (P< 0.05). Conclusion Epinephrine combined with sodium hydrogen exchanger isoform 1 inhibitor Cariporide may represent a novel and remarkably effective intervention for resuscitation from prolonged VF.

System integration of China's first PEMFC locomotive

In the face of growing environmental pollution, developing a fuel-cell-driven shunting locomotive is a great challenge in China for environmental protection and energy saving, which combines the environmental advantages of an electric locomotive with the lower infrastructure costs of a diesel-electric locomotive. In this paper, the investigation status and the development trend of the fuel-cell-driven shunting locomotive were introduced. Through innovation of the power system using fuel cells, an experiment prototype of a fuel-cell shunting locomotive was developed, which would reduce the effects on the environment of the existing locomotives. This was the first locomotive to use a proton exchange membrane fuel-cell （PEMFC） power plant in China. From October 2012, we started to test the fuel-cell power plant and further test runs on the test rail-line in Chengdu, Sichuan. The achieved encouraging results can provide fundamental data for the modification of the current individual fuel cell locomotives or further development of the fuel-cell hybrid ones in China.

Improvement of hydrogen isotope exchange reactions on Li_4SiO_4 ceramic pebble by catalytic metals

Li4SiO4 ceramic pebble is considered as a candidate tritium breeding material of Chinese Helium Cooled Solid Breeder Test Blanket Module （CH HCSB TBM） for the International Thermonuclear Experimental Reactor （ITER）. In this paper, LiaSiO4 ceramic pebbles deposited with catalytic metals, including Pt, Pd, Ru and Ir, were prepared by wet impregnation method. The metal particles on Li4SiO4 pebble exhibit a good promotion of hydrogen isotope exchange reactions in H2-D20 gas system, with conversion equilibrium temperature reduction of 200-300 ~C. The out-of-pile tritium release experiments were performed using 1.0 wt% Pt/Li4SiO4 and Li4SiO4 pebbles irradiated in a thermal neutron reactor. The thermal desorption spectroscopy shows that Pt was effective to increase the tritium release rate at lower temperatures, and the ratio of tritium molecule （HT） to tritiated water （HTO） of 1.0 wt% Pt/LiaSiO4 was much more than that of Li4SiO4, which released mainly as HTO. Thus, catalytic metals deposited on LiaSiO4 pebble may help to accelerate the recovery of bred tritium particularly in low temperature region, and increase the tritium molecule form released from the tritium breedin~ materials.

Highly active iridium catalyst for hydrogen production from formic acid

Formic acid（FA） dehydrogenation has attracted a lot of attentions since it is a convenient method for H_2 production. In this work, we designed a self-supporting fuel cell system, in which H_2 from FA is supplied into the fuel cell, and the exhaust heat from the fuel cell supported the FA dehydrogenation. In order to realize the system, we synthesized a highly active and selective homogeneous catalyst Ir Cp＊Cl_2 bpym for FA dehydrogenation. The turnover frequency（TOF） of the catalyst for FA dehydrogenation is as high as7150 h^（-1）at 50°C, and is up to 144,000 h^（-1）at 90°C. The catalyst also shows excellent catalytic stability for FA dehydrogenation after several cycles of test. The conversion ratio of FA can achieve 93.2%, and no carbon monoxide is detected in the evolved gas. Therefore, the evolved gas could be applied in the proton exchange membrane fuel cell（PEMFC） directly. This is a potential technology for hydrogen storage and generation. The power density of the PEMFC driven by the evolved gas could approximate to that using pure hydrogen.

Visible Light Promoted Direct Deuteration of Alkenes via Co(III)–H Mediated H/D Exchange

We report herein a visible light-mediated direct deuteration of alkenes with D_(2)O or deuterated methanol(MeOD)using a cobaloxime as a hydrogen/deuterium(H/D)exchange catalyst.The synergistic photoredox/Co catalysis enabled facile deuterium(D)-incorporation of a variety of terminal and internal alkenes at either terminal or benzylic positions.We proposed that this process proceeded through a sequence of reversible addition-elimination reactions and fast proton exchange involving Co(III)–H,which was generated in situ by photoreduction.

Subfunctionalization of a monolignol to a phytoalexin glucosyltransferase is accompanied by substrate inhibition

Uridine diphosphate-dependent glycosyltransferases(UGTs)mediate the glycosylation of plant metabolites,thereby altering their physicochemical properties and bioactivities.Plants possess numerous UGT genes,with the encoded enzymes often glycosylating multiple substrates and some exhibiting substrate inhibition kinetics,but the biological function and molecular basis of these phenomena are not fully understood.The promiscuous monolignol/phytoalexin glycosyltransferase NbUGT72AY1 exhibits substrate inhibition(Ki)at 4 mM scopoletin,whereas the highly homologous monolignol StUGT72AY2 is inhibited at 190 mM.We therefore used hydrogen/deuterium exchange mass spectrometry and structure-based mutational analyses of both proteins and introduced NbUGT72AY1 residues into StUGT72AY2 and vice versa to study promiscuity and substrate inhibition of UGTs.A single F87I and chimeric mutant of NbUGT72AY1 showed significantly reducedscopoletin substrate inhibition,whereas its monolignolgly cosylation activity was almost unaffected.Reverse mutations in StUGT72AY2 resulted in increased scopoletin glycosylation,leading to enhanced promiscuity,which was accompanied by substrate inhibition.Studies of 3D structures identified open and closed UGT conformers,allowing visualization of the dynamics of conformational changes that occur during catalysis.Previously postulated substrate access tunnels likely serve as drainage channels.The results suggest a two-site model in which the second substrate molecule binds near the catalytic site and blocks product release.Mutational studies showed that minor changes in amino acid sequence can enhance the promiscuity of the enzyme and add new capabilities such as substrate inhibition without affecting existing functions.The proposed subfunctionalization mechanism of expanded promiscuity may play a role in enzyme evolution and highlights the importance of promiscuous enzymes in providing new functions.

Clean SEA-TROSY Experiments to Map Solvent Exposed Amides in Large Proteins

It is well known that the SEA-TROSY experiment could alleviate some of the problems of resonance overlap in 15N/2H labeled proteins as it was designed to selectively map solvent exposed amide protons. However, SEA- TROSY spectra may be contaminated with exchange-relayed NOE contributions from fast exchanged hydroxyl or amine protons and contributions from longitudinal relaxation. Also, perdeuteration of the protein sample is a pre-requisite for this experiment. In this communication, a modified version, clean SEA-TROSY, was proposed to eliminate these artifacts and to allow the experiment to be applied to protonated or partially deuterated proteins and protein complexes.

Study of Structural Stability of Cyclophilin A by NMR and Circular Dichroism Spectra

The structural stability of cyclophilin A （CypA） was investigated using H/D exchange and temperature coefficients of chemical shifts of amide protons, monitored by 213 heteronuclear NMR spectroscopy. Amide proton exchange rates were measured by H/D exchange experiments for slow-exchange protons and measured by SEA （Solvent Exposed Amides）-HSQC experiments for fast-exchange protons. Temperature coefficients of chemical shifts and hydrogen exchange rates of amide protons show reasonably good correlation with the protein structure. Totally, 44 out of 153 non-proline assigned residues still exist in 86 d of hydrogen-deuterium exchange at 4 ℃, suggesting that CypA structure should be highly stable. Residues in secondary structures of α2, β1, β2, β5, β6 and β7 might constitute the hydrophobic core of the protein. The change in free energy of unfolding （ △Gu^H2O ） of CypA was estimated to be （21.99± 1.53） kJ·mol^-1 by circular dichroism （CD）. The large free energy change is also an indicator of the high structural stability.