Direct structure determination using residual dipolar couplings: reaction-site conformation of methionine sulfoxide reductase in solution

Residual dipolar couplings (RDC) from partially aligned molecules provide long-range structural data and are thus particularly well adapted to rapid structure validation or protein fold recognition. Extensive measurements in two alignment media can also provide precise de novo structure from RDC alone. We have applied a novel combination of these approaches to the study of methionine sulfoxide reductase (MsrA) from Erwinia chrysanthemi, a 27 kDa enzyme essential for repairing oxidative stress damage. The tertiary fold was initially validated by comparing backbone RDC to expected values from the crystal structure of the homologous MsrA from Escherichia coli. Good agreement was found throughout the chain, verifying the overall topology of the molecule, with the exception of the catalytically important peptide P196-L202, where strong and systematic RDC violation was observed. No evidence for local differential mobility in this region was detected, implying that the structure of the strand differs in the two molecules. We have therefore applied the de novo approach meccano to determine the conformation of this peptide using only RDC. A single conformation is found that is in agreement with all measured data. The aligned peptide can be docked onto the expected covalence of the rest of the template molecule while respecting its strictly defined relative orientation. In contrast to the structure of MsrA from E. coli, the reactive side chain of Cys200 is oriented toward the interior of the molecule and therefore closer to the catalytic Cys53, obviating the need for previously proposed conformational reorganization prior to formation of this disulfide intermediate. This analysis requires only backbone assignment and uses unambiguously assigned and readily measurable structural data, thereby greatly economizing investigation time compared to established nuclear Overhauser effect- (nOe-) based structure calculation methods.     

An easy-to-use estimate of the energy-release rate for crack arrays

Formation of crack arrays plays an increasing role in several fields of applied physics. The energy-release rate of the cracks controls the development of the array. Therefore, following the concept of configurational forces, a simplified analytical expression is provided for the energy-release rate, which is based both on numerical studies and on a specially adapted beam model. Comparisons of this easy-to-use estimate of the energy-release rate with established results from the literature as well as detailed numerical results are presented. The provided estimate of the energy-release rate can easily be extended to non-equidistant cracks and an anisotropic material.     

Fluorotrimethyl[(Z)-pentafluoropropen-1-yl]phosphorane: Structure,Bonding, and Reactivity

In this contribution we report on fluorotrimethyl[(Z)-pentafluoropropen-1-yl]phosphorane as a phosphorus based fluorinating reagent. Its solid state structure can be described as a trigonal bipyramid featuring elongated axial bonds due to the formation of a 3-center 4-electron bond. Abstraction of the fluoride ion leads to a shortening of the axial P–C bond. Thus the title compound can be utilized for substitution of bromine with fluorine and for the transfer of fluoride ions onto electrophilic compounds. Reaction with Sn(C₂F₅)₂Br₂ afforded salt [P(CH₃)₃(C₃F₅)]₂[Sn(C₂F₅)₂F₄]. When fluorotrimethyl[(Z)-pentafluoropropen-1-yl]phosphorane was treated with P(C₂F₅)₂F the primarily produced anion is sufficiently nucleophilic to attack the propenyl group of the cation in β-position to the phosphorus atom to yield zwitterionic [Me₃PCF=C(CF₃)–PF₃(C₂F₅)₂].     

Crystalline Divinyldiarsenes and Cleavage of the As=As Bond

The first divinyldiarsenes [{(NHC)C(Ph)}As]₂ (NHC=IPr 3 a , SIPr 3 b ; IPr=C{(NAr)CH}₂; SIPr=C{(NAr)CH₂}₂; Ar=2,6-iPr₂C₆H₃) are reported. Compounds 3 a and 3 b were prepared by the reduction of corresponding chlorides {(NHC)C(Ph)}AsCl₂ (NHC=IPr 2 a , SIPr 2 b ) with Mg. Calculations revealed a small HOMO–LUMO energy gap of 3.86 ( 3 a ) and 4.24 eV ( 3 b ). Treatment of 3 a with (Me₂S)AuCl led to the cleavage of the As=As bond to restore 2 a , which is expected to proceed via the diarsane [{(IPr)C(Ph)}AsCl]₂ ( 4 ). Remarkably, 4 as well as 2 a can be selectively accessed on treatment of 3 a with an appropriate amount of C₂Cl₆. Moreover, 3 a readily reacts with PhEEPh (E=Se or Te) at room temperature to give {(IPr)C(Ph)}As(EPh)₂ (E=Se 5 a ; Te 5 b ), revealing the cleavage of As=As and E−E bonds and the formation of As−E bonds. Such highly selective stepwise oxidation ( 3 a → 4 → 2 a ) and bond metathesis ( 3 a → 5 a , b ) reactions are unprecedented in main-group chemistry.     

Differential Response of Pentanal and Hexanal Exhalation to Supplemental Oxygen and Mechanical Ventilation in Rats

High inspired oxygen during mechanical ventilation may influence the exhalation of the previously proposed breath biomarkers pentanal and hexanal, and additionally induce systemic inflammation. We therefore investigated the effect of various concentrations of inspired oxygen on pentanal and hexanal exhalation and serum interleukin concentrations in 30 Sprague Dawley rats mechanically ventilated with 30, 60, or 93% inspired oxygen for 12 h. Pentanal exhalation did not differ as a function of inspired oxygen but increased by an average of 0.4 (95%CI: 0.3; 0.5) ppb per hour, with concentrations doubling from 3.8 (IQR: 2.8; 5.1) ppb at baseline to 7.3 (IQR: 5.0; 10.8) ppb after 12 h. Hexanal exhalation was slightly higher at 93% of inspired oxygen with an average difference of 0.09 (95%CI: 0.002; 0.172) ppb compared to 30%. Serum IL-6 did not differ by inspired oxygen, whereas IL-10 at 60% and 93% of inspired oxygen was greater than with 30%. Both interleukins increased over 12 h of mechanical ventilation at all oxygen concentrations. Mechanical ventilation at high inspired oxygen promotes pulmonary lipid peroxidation and systemic inflammation. However, the response of pentanal and hexanal exhalation varies, with pentanal increasing by mechanical ventilation, whereas hexanal increases by high inspired oxygen concentrations.     

Exploring the Reactivity of a Frustrated Sn/P Lewis Pair: The Highly Selective Complexation of the cis-Azobenzene Photoisomer

The reactivity of the geminal frustrated Lewis pair (FLP) (F₅C₂)₃SnCH₂P(tBu)₂ ( 1 ) was explored by reacting it with a variety of small molecules (PhOCN, PhNCS, PhCCH, tBuCCH, H₃CC(O)CH=CH₂, Ph[C(O)]₂Ph, PhN=NPh and Me₃SiCHN₂), featuring polar or non-polar multiple bonds and/or represent α,β-unsaturated systems. While most adducts are formed readily, the binding of azobenzene requires UV-induced photoisomerization, which results in the highly selective complexation of cis-azobenzene. In the case of benzil, the reaction does not lead to the expected 1,2- or 1,4-addition products, but to the non-stereoselective (tBu)₂PCH₂-transfer to a prochiral keto function of benzil. All adducts of 1 were characterised by means of multinuclear NMR spectroscopy, elemental analyses and X-ray diffraction experiments.     

Triphenylphosphine-Catalyzed Isomerisation of Propargyl Bromide to 1-Bromopropadiene

Pure bromopropadiene, BrCH=C=CH₂ has been obtained by isomerisation of propargyl bromide in the presence of catalytic amounts of triphenylphosphine.     

Mesoionic Dithiolates (MIDts) Derived from 1,3-Imidazole-Based Anionic Dicarbenes (ADCs)

Mesoionic dithiolates [(MIDt^Ar)Li(LiBr)₂(THF)₃] (MIDt^Ar={SC(NDipp)}₂CAr; Dipp=2,6-iPr₂C₆H₃; Ar=Ph 3 a , 3-MeC₆H₄ (3-Tol) 3 b , 4-Me₂NC₆H₄ (DMP) 3 c ) and [(MIDt^Ph)Li(THF)₂] ( 4 ) are readily accessible (in≥90 % yields) as crystalline solids on treatments of anionic dicarbenes Li(ADC^Ar) ( 2 a - c ) (ADC^Ar={C(NDipp)₂}₂CAr) with elemental sulfur. 3 a - c and 4 are monoanionic ditopic ligands with both the sulfur atoms formally negatively charged, while the 1,3-imidazole unit bears a formal positive charge. Treatment of 4 with (L)GeCl₂ (L=1,4-dioxane) affords the germylene (MIDt^Ph)GeCl ( 5 ) featuring a three-coordinated Ge atom. 5 reacts with (L)GeCl₂ to give the Ge−Ge catenation product (MIDt^Ph)GeGeCl₃ ( 6 ). KC₈ reduction of 5 yields the homoleptic germylene (MIDt^Ph)₂Ge ( 7 ). Compounds 3 a - c and 4 – 7 have been characterized by spectroscopic studies and single-crystal X-ray diffraction. The electronic structures of 4 – 7 have been analyzed by DFT calculations.     

Bidentate Lewis Acids Derived from o-Diethynylbenzene with Group 13 and 14 Functions

Starting from 1,2-diethynylbenzene, a series of bidentate Lewis acids was prepared by means of hydrometalations, in particular hydrosilylation, hydroboration, hydroalumination and terminal metalation based on group 13 and 14 elements. In the case of terminal alkyne metalation, the Lewis-acidic gallium function was introduced using triethylgallium under alkane elimination. A total of six different Lewis acids based on a semiflexible organic scaffold were prepared, bearing −SiClMe₂, −SiCl₂Me, −SiCl₃, −B(C₆F₅)₂, −AlBis₂ (Bis=bis(trimethylsilyl)methyl) and −GaEt₂ as the corresponding functional units. In all cases, the Lewis acid functionalisation was carried out twice and the products were obtained in good to excellent yields. In the case of the twofold gallium Lewis acid, a different structural motif in the form of a polymer-like chain was observed in the solid state. All new bidentate Lewis acids were characterised by multinuclear NMR spectroscopy, CHN analysis and X-ray diffraction experiments.