Einwirkung von Schwefel und Ammoniak auf Methyl-neopentylketon

Zusammenfassung Bei der Einwirkung von Schwefel und Ammoniak auf Methylneopentylketon in Pyridin bei Raumtemp. bilden sich die beiden theoretisch möglichen Thiazoline-³, das 2,4-Dimethyl-2-neopentyl-5-tert. butyl-thiazolin-³ und das 2-Methyl-2,4-di-neopentyl-thiazolin-³ (etwa im Verhältnis 793). Daneben entsteht 2-Methyl-2,4-dineopentyl-imidazolin-³. Die Darstellung und Charakterisierung der sich vom Methylneopentylketon ableitenden reinen isomeren -Brom- bzw. -Mercaptoketone sowie einiger daraus synthetisierter Thiazoline-³ bzw. Thiazole wird beschrieben.Für Korrespondenz ist diese Adresse zu verwenden.49. Mitt.:F. Asinger, W. Schäfer undA. Saus, Mh. Chem.96, 1278 (1965).Teil der DissertationF. Gentz, Techn. Univ. Dresden 1963.     

Quantum dynamics of H2 formation on a graphite surface through the Langmuir Hinshelwood mechanism

We have studied the formation of the H2 molecule on a graphite surface, when both H atoms are initially physisorbed. The graphite surface is assumed to be planar, and a model potential is obtained in a semiempirical way to reproduce the experimental properties of H physisorption on graphite. The reaction probability has been computed in the case when the angular momentum of the relative H-H motion lies parallel to the surface plane. Three-dimensional wave packet calculations have been performed for collision energies ranging from 2 to 50 meV. It is shown that the reaction occurs with a significant probability and produces the H2 molecule with a considerable amount of vibrationnal energy. A simple mechanical model is presented, where desorption of the nascent H2 molecule results from two successive binary elastic collisions.     

Solution structure, mutagenesis, and NH exchange studies of the MutT enzyme–Mg-8-oxo-dGMP complex

The MutT pyrophosphohydrolase from E. coli (129 residues) catalyzes the hydrolysis of nucleoside triphosphates (NTP), including 8-oxo-dGTP, by substitution at Pβ, to yield NMP and pyrophosphate. The product, 8-oxo-dGMP is an unusually tight binding, slowly exchanging inhibitor with a K_D=52 nM, (ΔG°=−9.8 kcal/mol) which is 6.1 kcal/mol tighter than the binding of dGMP (ΔG°=−3.7 kcal/mol). The higher affinity for 8-oxo-dGMP results from a more favorable ΔH_binding (−32 kcal/mol) despite an unfavorable −TΔS°_binding (+22 kcal/mol). The solution structure of the MutT–Mg²⁺-8-oxo-dGMP complex shows a narrowed, hydrophobic nucleotide-binding cleft with Asn-119 and Arg-78 among the few polar residues. The N119A, N119D, R78K and R78A single mutations, and the R78K+N119A double mutant all showed largely intact active sites, on the basis of small changes in the kinetic parameters of dGTP hydrolysis and in ¹H–¹⁵N HSQC spectra. However, the N119A mutation profoundly weakened the active site binding of 8-oxo-dGMP by 4.3 kcal/mol (1650-fold). The N119D mutation also weakened 8-oxo-dGMP binding but only by 2.1 kcal/mol (37-fold), suggesting that Asn-119 functioned both as a hydrogen bond donor to C8=O, and a hydrogen bond acceptor from N7H of 8-oxo-dGMP, while aspartate at position −119 functioned as an acceptor of a single hydrogen bond. Much smaller weakening effects (0.3–0.4 kcal/mol) on the binding of dGMP and dAMP were found, indicating specific hydrogen bonding of Asn-119 to 8-oxo-dGMP. While formation of the wild type MutT–Mg²⁺-8-oxo-dGMP complex slowed the backbone NH exchange rates of 45 residues distributed throughout the protein, the same complex of the N119A mutant slowed the exchange rates of only 11 residues at or near the active site, indicating an increase in conformational flexibility of the N119A mutant. The R78K and R78A mutations weakened the binding of 8-oxo-dGMP by 1.7 and 1.1 kcal/mol, respectively, indicating a lesser role of Arg-78 than of Asn-119 in the selective binding of 8-oxo-dGMP, likely donating a single hydrogen bond to its C6=O. The R78K+N119A double mutant weakened the binding of 8-oxo-dGMP (K_I^slope=3.1 mM) by 6.5±0.2 kcal/mol which overlaps, within error with the sum of the effects of the two single mutants (6.0±0.3 kcal/mol). Such additive effects of the two single mutants in the double mutant are most simply explained by the independent functioning of Asn-119 and Arg-78 in the binding of 8-oxo-dGMP. Independent functioning of these two residues in nucleotide binding is consistent with their locations in the MutT–Mg²⁺-8-oxo-dGMP complex, on opposite sides of the active site cleft, with a distance of 8.4±0.5 Å between their side chain nitrogens.     

Simultaneous separation and identification of hashish constituents by coupled liquid chromatography-mass spectrometry (HPLC-MS)

Summary This paper reports the use of liquid chromatography for the separation and determination of the major cannabinoids extracted from hashish samples. The direct coupling to the mass spectrometer enables the selective identification both of neutral and acidic cannabinoids. The developed method does not require any preliminary derivatization and should, therefore, be of interest in forensic analysis for simple and unequivocal determination of hashish constituents.Part of this work was presented at the 2nd Italian-Spanish Joint Meeting of Medicinal Chemistry, 30 August–3 September 1995, Ferrara, Italy.     

Peptide labeling using 188Re, 188Re-MAG3 and 153Sm-H1ETA: A comparison on their in vitro lipophilicity

Lanreotide peptide was labeled with ¹⁵³Sm-H₁ETA and ¹⁸⁸Re-MAG₃ in order to evaluate whether or not their conjugation to the peptide produce significant differences of the in vitro lipophilicity with respect to the ¹⁸⁸Re-lanreotide prepared by the direct labeling method (highly lipophilic). The differences of lipophilicity between the complexes, were evaluated using a reverse phase HPLC system. The measured lipophilicity of ¹⁵³Sm-H₁ETA-lanreotide, ¹⁸⁸Re-MAG₃-lanreotide and ¹⁸⁸Re-lanreotide was taken to be the capacity factor [k" = (t _R-t ₀)/t ₀ where t _R is the retention time and t ₀ is the dead time] for each of the complexes under identical chromatography conditions. Results showed that the in vitro lipophilicity decreased in the order ¹⁸⁸Re-lanreotide (direct labeling), ¹⁸⁸Re-MAG₃-lanreotide and ¹⁵³Sm-H₁ETA-lanreotide. Since the last one has a capacity factor (k") similar to that of ¹⁸⁸Re-MAG₃, some renal elimination for ¹⁵³Sm-H₁ETA-lanreotide could be expected, which probably would reduce the unnecessary radiation dose to normal tissues.     

Mixed C18 and C1 modification on an optical fiber for chromatographic sensing

An optical fiber-chromatographic sensor, aiming at simultaneous and selective response to multiple components following a chromatographic separation, is described. We report an improved approach for immobilization of octadecyl (C(18)) and methyl (C(1)) moieties as stationary phase on an optical fiber suitable as a sensing phase for organic solutes. By this approach, the stability and lifetime of the sensing layer as well as the detectability and retention behavior of the chromatographic sensor could be improved. Infrared spectroscopy was employed to confirm the presence of C(18) and C(1) moieties on the modified surface of the optical fiber. The chromatographic sensor was applied, with good sensitivity and chemical selectivity, to the simultaneous separation and detection of bromobenzene and toluene, using water as the mobile phase.     

Nanoscale organization of the pathogen receptor DC-SIGN mapped by single-molecule high-resolution fluorescence microscopy.

DC-SIGN, a C-type lectin exclusively expressed on dendritic cells (DCs), plays an important role in pathogen recognition by binding with high affinity to a large variety of microorganisms. Recent experimental evidence points to a direct relation between the function of DC-SIGN as a viral receptor and its spatial arrangement on the plasma membrane. We have investigated the nanoscale organization of fluorescently labeled DC-SIGN on intact isolated DCs by means of near-field scanning optical microscopy (NSOM) combined with single-molecule detection. Fluorescence spots of different intensity and size have been directly visualized by optical means with a spatial resolution of less than 100 nm. Intensity- and size-distribution histograms of the DC-SIGN fluorescent spots confirm that approximately 80 % of the receptors are organized in nanosized domains randomly distributed on the cell membrane. Intensity-size correlation analysis revealed remarkable heterogeneity in the molecular packing density of the domains. Furthermore, we have mapped the intermolecular organization within a dense cluster by means of sequential NSOM imaging combined with discrete single-molecule photobleaching. In this way we have determined the spatial coordinates of 13 different individual dyes, with a localization accuracy of 6 nm. Our experimental observations are all consistent with an arrangement of DC-SIGN designed to maximize its chances of binding to a wide range of microorganisms. Our data also illustrate the potential of NSOM as an ultrasensitive, high-resolution technique to probe nanometer-scale organization of molecules on the cell membrane.     

Preparation and electrochemical behavior of ordered rh adlayers on Pt(100) electrodes

Rhodium adlayers on Pt(100) substrates have been prepared by electrodeposition from dilute Rh(III) acidic solutions. The initially disordered layer is electrochemically annealed by applying a polarization program consisting of high-sweep-rate multicycle sequences between 0.05 and 0.78 V(RHE) in 0.1 M H(2)SO(4). In this way, a pseudomorphic Rh monolayer can be prepared on Pt(100) substrates. The degree of order of the electrochemically annealed layer has been evidenced not only through voltammetric experiments but also by means of scanning tunneling microscopy with atomic resolution for iodine-protected adlayers, which show a c(2 x 2) structure. The electrochemically induced ordering of the Rh adlayer appears to be a consequence of the repeated cycles of adsorption/desorption of H and, especially, oxygenated species. Voltammetry in sulfuric acid solutions permits examination of the energetics of H/anions and OH/O adsorption as a function of the Rh coverage. The first monolayer adsorbs both hydrogen and oxygenated species more strongly than the second one. This can be explained through an electronic effect caused by the underlying Pt(100) substrate.