Nuclear resonance vibrational spectroscopy reveals the FeS cluster composition and active site vibrational properties of an O2-tolerant NAD+-reducing [NiFe] hydrogenase

Hydrogenases are complex metalloenzymes that catalyze the reversible splitting of molecular hydrogen into protons and electrons essentially without overpotential. The NAD⁺-reducing soluble hydrogenase (SH) from Ralstonia eutropha is capable of H₂ conversion even in the presence of usually toxic dioxygen. The molecular details of the underlying reactions are largely unknown, mainly because of limited knowledge of the structure and function of the various metal cofactors present in the enzyme. Here, all iron-containing cofactors of the SH were investigated by ⁵⁷Fe specific nuclear resonance vibrational spectroscopy (NRVS). Our data provide experimental evidence for one [2Fe2S] center and four [4Fe4S] clusters, which is consistent with the amino acid sequence composition. Only the [2Fe2S] cluster and one of the four [4Fe4S] clusters were reduced upon incubation of the SH with NADH. This finding explains the discrepancy between the large number of FeS clusters and the small amount of FeS cluster-related signals as detected by electron paramagnetic resonance spectroscopic analysis of several NAD⁺-reducing hydrogenases. For the first time, Fe–CO and Fe–CN modes derived from the [NiFe] active site could be distinguished by NRVS through selective ¹³C labeling of the CO ligand. This strategy also revealed the molecular coordinates that dominate the individual Fe–CO modes. The present approach explores the complex vibrational signature of the Fe–S clusters and the hydrogenase active site, thereby showing that NRVS represents a powerful tool for the elucidation of complex biocatalysts containing multiple cofactors.     

Synthetic hydrogels as scaffolds for manipulating endothelium cell behaviors

Synthetic hydrogels can be used as scaffolds that not only favor endothelial cells（ECs） proliferation but also manipulate the behaviors and functions of the ECs.In this review paper,the effect of chemical structure,Young’s modulus （E） and zeta potential（ξ） of synthetic hydrogel scaffolds on static cell behaviors,including cell morphology,proliferation, cytoskeleton structure and focal adhesion,and on dynamic cell behaviors,including migration velocity and morphology oscillation,as well as on EC function such as anti-platelet adhesion,are reported.It was found that negatively charged hydrogels,poly（2-acrylamido-2-methylpropanesulfonic sodium）（PNaAMPS） and poly（sodium p-styrene sulphonate） （PNaSS）,can directly promote cell proliferation,with no need of surface modification by any cell-adhesive proteins or peptides at the environment of serum-containing medium.In addition,the Young’s modulus（E） and zeta potential（ξ） of hydrogel scaffolds are quantitatively tuned by copolymer hydrogels,poly（NaAMPS-co-DMAAm） and poly（NaSS-co-DMAAm）, in which the two kinds of negatively charged monomers NaAMPS and NaSS are copolymerized with neutral monomer,N,N-dimethylacrylamide（DMAAm）.It was found that the critical zeta potential of hydrogels manipulating EC morphology,proliferation,and motility isξ_critical= -20.83 mV andξ_critical= -14.0 mV for poly（NaAMPS-co-DMAAm） and poly（NaSS-co-DMAAm）,respectively.The above mentioned EC behaviors well correlate with the adsorption of fibronectin, a kind of cell-adhesive protein,on the hydrogel surfaces.Furthermore,adhered platelets on the EC monolayers cultured on the hydrogel scaffolds obviously decreases with an increase of the Young’s modulus（E） of the hydrogels,especially when E>60 kPa.Glycocalyx assay and gene expression of ECs demonstrate that the anti-platelet adhesion well correlates with the EC-specific glycocalyx.The above investigation suggests that understanding the relationship between physic-chemical properties of synthetic hydrogels and cell responses is essential to design optimal soft and wet scaffolds for tissue engineering.     

Oxo-tethered ruthenium(II) complex as a bifunctional catalyst for asymmetric transfer hydrogenation and H2 hydrogenation

Newly developed oxo-tethered Ru amido complexes (R,R)-1 and their HCl adducts (R,R)-2 exhibited excellent catalytic performance for both asymmetric transfer hydrogenation and the hydrogenation of ketonic substrates under neutral conditions without any cocatalysts to give chiral secondary alcohols with high levels of enantioselectivity.     

Effects of Temperature and Mobile Phase Condition on Chiral Recognition of Poly(l-phenylalanine) Chiral Stationary Phase

Characteristics of the chiral stationary phase with poly(l-phenylalanine) peptide selector, which was in ??-helical state, was reported. Since environmental factors affect peptide conformation, the changes in enantioselectivity were examined depending on column temperature and mobile phase conditions (ionic strength, pH, mobile phase composition). Column temperature and pH drastically affected the enantioselectivity. Based on these changes, the relation between chiral recognition and secondary structure of the peptide selector was discussed. The column stability during sequential analysis under different separation conditions was also evaluated.     

Synthesis of oligonucleotides possessing versatile probes for PET labelling and their rapid ligand-free click reaction

We synthesised aryl acetylene derivatives as versatile probes for labelling of oligonucleotides. RNA oligomers bearing an aryl acetylene molecule rapidly reacted with benzylazide derivatives under ligand-free click reaction conditions.     

Deformation and molecular orientation in films of metallocene polypropylene,ethylene–butylene rubber,and their 80/20 (w/w) blend revealed by the simultaneous kinetic measurement of microscopic infrared dichroism,macroscopic stress,and mesoscale strain

Simultaneous kinetic measurement of microscopic infrared dichroism, macroscopic stress, and mesoscale strain was used to study the deformation mechanisms of metallocene polypropylene (MPP), ethylene–butylene rubber (EBR), and their blend (MPP/EBR = 80/20 w/w). As with pure MPP, the molecular orientation in the blend is dominated by the necking of the isotactic polypropylene matrix. During the necking passage through the mesoscale sampling area, the molecular orientation of the polypropylene matrix in the blend is smaller than that in the pure polypropylene film at the same level of mesoscale strain. However, the orientation of the EBR dispersed phase in the blend is larger than that in the pure EBR film. This may result from the partial miscibility of the two ingredients in the amorphous phases and their resultant strong interfacial interaction. The large stress supported by the MPP matrix extends to the island of the EBR domain and leads to its large deformation. © 2005 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 43: 1520–1531, 2005     

Oxygenation of alkyl sulfides with ferrous perchlorate/ascorbic acid/oxygen system

In the oxidation of alkyl sulfides bearing acidic α-hydrogens eiher with phenobarbital induced rabbit liver microsomal cytochrome P-450, or with the Udenfriend's model system (ferous perchlorate/ascorbic acid/oxygen system), both S-dealkylation and S-monoxygenation took place concurrently. Meanwhile, in the oxidation of simple alkyl sulfides, steeoselective S-monooxygenation was found of occur predominantly.