Synthesis and bulk characterization of new P(CB‐b‐S) diblock copolymers

Strongly asymmetric chlorinated polybutadiene‐b‐polystyrene, [P((CB)_x‐b‐(PS)_y)] diblock copolymers with increasing x/(x + y) ratios (up to 5.2 mol %) have been synthesized by the selective chlorination of the polybutadiene (PB) block in solution. Chlorination has been performed in anhydrous dichloromethane added with an antioxidant [2,2′‐methylenebis‐(6‐tert‐butyl‐4‐methyl‐phenol)], at −50°C, under a continuous Ar flow and in the dark. Under the optimized experimental conditions, the PB chlorination is not complete, but the PS block is left unmodified. Even in the presence of a large chlorine excess (Cl₂/butene unit molar ratio of 2.5), the experimental degree of chlorination of homo PB does not exceed 85%. The chlorinated copolymers have been characterized by ¹H‐NMR, IR spectroscopy, size‐exclusion chromatography, and elemental analysis. The chlorinated copolymers have also been studied by DSC and SAXS after annealing at 150°C. Although at this temperature the parent homopolymers are immiscible, no microphase separation has been observed for the block copolymers. © 1999 John Wiley & Sons, Inc. J Polym Sci A: Polym Chem 37: 233–244, 1999     

Assignments of the Pfr-Pr FTIR difference spectrum of cyanobacterial phytochrome Cph1 using 15N and 13C isotopically labeled phycocyanobilin chromophore

The reversible red and far-red light-induced transitions of cyanobacterial phytochrome Cph1 from Synechocystis PCC 6803 were investigated by Fourier transform infrared (FTIR) difference spectroscopy. High-quality light-induced Pfr-Pr difference FTIR spectra were recorded for the 58 kDa N-terminal domain of Cph1 by repetitive photochemical cycling and signal averaging. The Pfr-Pr difference spectra in H(2)O and D(2)O were very similar to those previously reported for full-length 85 kDa Cph1.(1) Published assignments were extended by analysis of the effects of (13)C and (15)N isotope substitutions at selected sites in the phycocyanobilin chromophore and by (15)N global labeling of the protein. The Pfr-Pr difference spectra were dominated by an amide I peak/trough at 1653 cm(-1)(+)/1631 cm(-1)(-) and a smaller amide II band at 1554 cm(-1). Labeling effects allowed specific chromophore assignments for the C(1)=O (1736 cm(-1)(-)/1724 cm(-1)(+)) and C(19)=O (1704 cm(-1)(-)) carbonyl vibrations, C=C vibrations at 1589 cm(-1)(+), and bands at 1537(-), 1512(+), 1491(-), 1163(+), 1151(-), 1134(+), 1109(-), and 1072(-) cm(-1) that must involve chromophore C-N bonds. A variety of additional changes were insensitive to isotope labeling of the chromophore. Effects of (15)N labeling of the protein were used to tentatively assign some of these to specific amino acid changes. Those insensitive to (15)N labeling included a protonated aspartic or glutamic acid at 1734 cm(-1)(-)/1722 cm(-1)(+) and a cysteine at 2575 cm(-1)(+)/2557 cm(-1)(-). Bands sensitive to (15)N protein labeling at 1487 cm(-1)(+)/1502 cm(-1)(-) might arise from trytophan and bands at 1261 cm(-1)(+)/1244 cm(-1)(-) and 1107 cm(-1)(-)/1095 cm(-1)(+) might arise from a histidine environment or protonation change. These assignments are discussed in light of the 15Z-E photoisomerization model of phototransformation and the associated protein conformational changes.     

Can luminescent quantum dots be efficient energy acceptors with organic dye donors?

We assessed the ability of luminescent quantum dots (QDs) to function as energy acceptors in fluorescence resonance energy transfer (FRET) assays, with organic dyes serving as donors. Either AlexaFluor 488 or Cy3 dye was attached to maltose binding protein (MBP) and used with various QD acceptors. Steady-state and time-resolved fluorescence measurements showed no apparent FRET from dye to QD. We attribute these observations to the dominance of a fast radiative decay rate of the donor excitation relative to a slow FRET decay rate. This is due to the long exciton lifetime of the acceptor compared to that of the dye, combined with substantial QD direct excitation.     

New chiral anion mediated asymmetric chemistry

Chemical reactions and processes often involve chiral, yet racemic, cationic reagents, intermediates or products. To afford instead non racemic or enantiopure compounds, an asymmetric ion pairing of the cations with enantiopure anions can be considered--the counter ions behaving as asymmetric auxiliaries, ligands or reagents. Detailed herein is a short review of our approach towards gaining reliable and predictable control over stereoselective ion pairing phenomena through the synthesis and the use of novel configurationally stable hexacoordinated phosphate anions.     

Analysis of a Complex Gaseous Mixture by TG-MS and TG-FTIR

The thermal decomposition of sodium ethyl xanthate (SEX) was used to compare the techniques of pyrolysis-gas chromatography-mass spectrometry (py-GC-MS), thermogravimetry-Fourier transform infrared spectroscopy (TG-FTIR), and TG-MS. In the py-GC-MS analysis, SEX was pyrolysed at 400°C in an inert atmosphere. Major gases evolved were carbon disulfide, diethyl sulfide, ethanol, and carbonyl sulfide. The TG of SEX exhibited a sharp mass loss at 201°C (42.3%) and a gradual mass loss at 217-325°C (20.8 %). The MS spectra of the evolved gases were complex due to overlapping of molecular, isotope, and fragment ion signals. Using the MS in selected ion monitoring mode, the major gases evolved were found to be carbon disulfide and carbonyl sulfide. The FTIR spectra of the evolved gases displayed vibrational frequencies due to alkanes, carbonyls, carbonyl sulfide, and carbon disulfide. From the analyses it was concluded that py-GC-MS provided unambiguous gas identification. Interpretation of the MS results was reliant on the py-GC-MS results, and the FTIR data was limited to identifying gases with very characteristic vibration frequencies. This revised version was published online in July 2006 with corrections to the Cover Date.     

N-Terminal Selective C−H Azidation of Proline-Containing Peptides: a Platform for Late-Stage Diversification

A methodology for the C−H azidation of N-terminal proline-containing peptides was developed employing only commercially available reagents. Peptides bearing a broad range of functionalities and containing up to 6 amino acids were selectively azidated at the carbamate-protected N-terminal residue in presence of the numerous other functional groups present on the molecules. Post-functionalizations of the obtained aminal compounds were achieved: cycloaddition reactions or C−C bond formations via a sequence of imine formation/nucleophilic addition were performed, offering an easy access to diversified peptides.     

The molecular structure of gaseous bis(hexamethydisilylamido)mercury(II), Hg{N(Si(CH3)3)2}2, as determined by electron diffraction

Gaseous bis(hexamethydisilylamido)mercury(II), Hg{N(SiMe₃)₂)₂}₂, has been studied by electron diffraction at a nozzle temperature of ca 390 K.

The diffraction data are consistent with a model consisting only of monomers. By assuming the NHgN chain to be linear and the HgHSi₂ fragments to be planar, an equilibrium conformer with a staggered Si₂NHgNSi₂ skeleton of D_ad-symmetry may be brought into a nice agreement with the observed diffraction data. The relatively large value of the vibrational amplitude of the inter-ligand SiSi distance, 0.26(12) A, indicates that the ligands undergo large amplitude vibrations about the NHgN axis. Steric considerations as well as the magnitude of the rotational barrier as estimated from the diffraction data (ca. 2 kcal mol⁻¹) show that this motion is hindered. A model with an eclipsed, co-planar Si₂NHgNSi₂ backbone of D_adsymmetry could not satisfactorily be brought into agreement with the observed diffraction data.

The values of some relevant key-parameters are: r_a(Hg---N) = 2.01(2) A, r_a(Si---N) = 1.732(9) A, r_a(Si---C) = 1.883(6) A;HgNSi = 116.0(1.0)°, SiNSi = 128.0(2.0)°, NSiC= 111.8(1.2)° and SiCH = 111.0(2.0)°. The trimethylsilyl groups are twisted 25(3)° away from their references positions typified by one Si---C bond of each such group eclipsing the adjacent Hg---N bond, in such a way that the overall symmetry of the model is lowered from D_ad to S₄.     

Hydrodynamic Rocking Disc Electrode Study of the TEMPO‐mediated Catalytic Oxidation of Primary Alcohols

The hydrodynamically thinned diffusion layer at a sinusoidally rocking disc is approximately uniform and can be expressed in terms of a diffusion layer thickness with D, the diffusion coefficient of the redox active species, v, the kinematic viscosity, Θ, the total rocking angle (here 90 degrees), and f, the rocking frequency (ranging here from 0.83 to 25 Hz). For the one‐electron oxidation of 2,2,6,6‐tetramethylpiperidine‐1‐oxyl (TEMPO) in aqueous carbonate buffer pH 9.5, it is shown that there is quantitative agreement between the expression for the diffusion layer thickness and experimental data. Next, for seven primary alcohols, the catalytic TEMPO‐mediated oxidation mechanism is investigated under rocking disc conditions. Chemical rate constants are evaluated (by varying the diffusion layer thickness) employing the DigiElch4.F simulation package. Trends in the chemical rate constants (compared with DFT calculations) suggest enhanced reactivity for methanol versus higher primary alcohols and for aromatic versus non‐aromatic primary alcohols. Rocking disc voltammetry allows quantitative mechanistic analysis in the laminar flow regime.     

Palladium Aryl Sulfonate Phosphine Catalysts for the Copolymerization of Acrylates with Ethene

The reaction of 2‐[bis(2‐methoxy‐phenyl)phosphanyl]‐4‐methyl‐benzenesulfonic acid (a) and 2‐[bis(2′,6′‐dimethoxybiphenyl‐2‐yl)phosphanyl]benzenesulfonic acid (b) with dimethyl(N,N,N′,N′‐tetramethylethylenediamine)‐palladium(II) (PdMe₂(TMEDA)) leads to the formation of TMEDA bridged palladium based polymerization catalysts ( 1a and 1b ). Upon reaction with pyridine, two mononuclear catalysts are formed ( 2a and 2b ). These catalysts are able to homopolymerize ethylene and also copolymerize ethylene with acrylates or with norbornenes. With ligand b , high molecular weight polymers are formed in high yields, but higher comonomer incorporations are obtained with ligand a .

     

Molecular structures and dynamics of the stepwise activation mechanism of a matrix metalloproteinase zymogen: challenging the cysteine switch dogma

Activation of matrix metalloproteinase zymogen (pro-MMP) is a vital homeostatic process, yet its molecular basis remains unresolved. Using stopped-flow X-ray spectroscopy of the active site zinc ion, we determined the temporal sequence of pro-MMP-9 activation catalyzed by tissue kallikrein protease in milliseconds to several minutes. The identity of three intermediates seen by X-ray spectroscopy was corroborated by molecular dynamics simulations and quantum mechanics/molecular mechanics calculations. The cysteine-zinc interaction that maintains enzyme latency is disrupted via active-site proton transfers that mediate transient metal-protein coordination events and eventual binding of water. Unexpectedly, these events ensue as a direct result of complexation of pro-MMP-9 and kallikrein and occur before proteolysis and eventual dissociation of the pro-peptide from the catalytic site. Here we demonstrate the synergism among long-range protein conformational transitions, local structural rearrangements, and fine atomic events in the process of zymogen activation.