Diels-alder cycloadditions of diene-substituted N-ethoxycarbonyl-2-methyl-1,2-dihydropyridines with N-phenylmaleimide

The ten possible substitution patterns for N-Ethoxycarbonyl-2-methyl-1,2-dihydropyridines 5 in which one or two olefinic sites are alkyl substituted were synthesized and reacted with N-phenylmaleimide 2 to provide cycloadducts 6. N-Ethoxycarbonyl-5,6-cyclohexyl-2-methyl-1,2-dihydropyridine 51 provided the novel spirocycle 61.     

The electrochemistry of benzo[b]quinolizinium ion

Summary In aprotic solvents, polarography and cyclic voltammetry indicate a two step reduction of acridizinium ion. The first reduction is a reversible one-electron step. The second involves two electrons and is irreversible. In situ ESR electrolysis at the potential of the first reduction gives no evidence of free radicals. Homogeneous chemical reduction of acridizinium by titration with sodium biphenylide indicates a one-electron reduction mechanism in solution. The absence of stable free radicals suggests a rapid post-reduction of the product.Addition of small amounts of water changes the aprotic solvent (mercury electrode) mechanism. The first wave becomes a two-electron irreversible wave at the expense of the second wave. In aqueous solvents two two-electron, irreversible reduction waves occur. Products of the second absorb and change the mechanism after the first CV scan.

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Elektrochemisches Verhalten des Benzo[b]chinolizinium-Ions

Zusammenfassung In aprotischen Lösungsmitteln wird durch Polarographie und cyclische Voltammetrie eine zweistufige Reduktion des Acridizinium-Ions angezeigt. Die erste Stufe ist reversibel und bezieht sich auf ein Elektron, die zweite Stufe umfaßt 2 Elektronen und ist irreversibel. Durch in situ ESR-Elektrolyse beim Potential der ersten Stufe konnte kein Beweis für freie Radikale gefunden werden. Homogene chemische Reduktion durch Titration mit Natriumbiphenylid deutet auf einen einelektronigen Reduktionsmechanismus hin. Durch Zugabe kleiner Wassermengen wird die erste Stufe auf Kosten der zweiten irreversibel und zweielektronig. In wäßrigen Lösungsmitteln treten 2 zweielektronige irreversible Reduktionsstufen auf.

     

Single-molecule tracking of sub-millisecond domain motion in calmodulin

We used single-pair fluorescence resonance energy transfer (spFRET) to track distance changes between domains of fluorescently labeled calmodulin (CaM) on the sub-millisecond time scale. In most cases, CaM remained in the same conformational substate over time periods of up to 1 ms, showing that conformational interchange occurs on a longer time scale. However, in some instances, apparent transitions between conformational substates could be detected. The magnitude of sub-millisecond motion within the dominant conformational substate also revealed fluctuations in distance between domains that were dependent on pH and ionic strength.     

Identification and confirmation of traces of chlorinated fatty acids in fish downstream of bleached kraft pulp mills by gas chromatography with halogen-specific detection

Methyl esters of threo-9,10-dichlorooctadecanoic, threo-7,8-dichlorohexadecanoic, and threo-5,6-dichlorotetradecanoic acids, present in transesterified extracts of filets, gonad, intestinal fat and carcass of white sucker (Catostomus commersoni) sampled in receiving waters of bleached kraft pulp mill effluents, were identified by gas chromatography with halogen-specific detection (XSD). Identification was based on (1) a comparison of the retention times of a sample peak with a prospective reference standard on two stationary phases of very different polarities by spiking, and (2) elution behavior of configurational and positional isomers of dichloro fatty acid methyl esters.     

Aliphatic-aromatic copolycarbonates derived from 2,2,4,4-tetramethyl-1,3-cyclobutanediol

The synthesis of copolycarbonates with high aliphatic contents for possible applications as optical plastics is described. Copolymers of 2,2,4,4-tetramethyl-1,3-cyclobutanediol (TMCBD) with a series of 14 bisphenol derivatives were prepared and characterized. The refractive index of these materials varies from 1.49 for the 2,2-hexafluoropropane substituted copolymer to 1.57 for the 9,9-fluorene substituted copolymer; thus, all of the copolymers exhibit lower refractive indices than the bisphenol A homopolycarbonate. The glass tran-sition temperatures of the copolymers vary from 110 to 210°C; the beta transition tem-peratures vary from -70 to 0°C when measured at 10 Hz by dynamic mechanical analysis. The copolymers exhibited good thermal stability (0–3% weight loss) when heated above the glass transition temperature, and only undergo extensive decomposition (80–90%) at 300–360°C. © 1995 John Wiley & Sons, Inc.     

Rapid characterization of soy protein and hydrolysates by capillary electrophoresis

Rapid methods for characterizing soy proteins and their hydrolysates by free-zone capillary electrophoresis were developed. The 7S- and 11S-rich protein fractions were separated by capillary electrophoresis (CE). The relationships between degrees of hydrolysis and electropherograms were established. Protein hydrolysates were separated in less than 6 min. Data showed that the CE method can be used effectively for monitoring protein hydrolysis during processing and for fingerprinting various types of protein products.     

Blue ferrocenium azurin: an organometalloprotein with tunable redox properties

A ferrocene derivative (2-[(methylsulfonyl)thio]ethylferrocene) (1) has been synthesized and incorporated into apo-azurin from Pseudomonas aeruginosa by covalent attachment to the highly conserved Cys112. The resulting artificial organometalloprotein (a protein containing organometallic compounds in the active site) has been characterized by UV-vis, electrospray mass spectrometry, and cyclic voltammetry (CV). Incorporation of 1 into azurin resulted in a higher solubility of the ferrocene group and improved stability of the ferrocenium species in aqueous solution, as shown by a more intense UV-vis absorption and a more reversible CV of the attached ferrocene group, respectively. The incorporation of 1 also increased the reduction potential of the complex from 402 to 579 mV (vs NHE), consistent with the ferrocene group being encapsulated inside the hydrophobic environment of the protein. Modulation of the reduction potential of ferrocene by residues near the secondary coordination sphere has also been demonstrated. Raising the pH from 4 to 9 resulted in a greater than 80 mV decrease in reduction potential of the protein-bound ferrocene (from 579 to 495 mV), while replacing Met121, an amino acid residue in close proximity to the ferrocene group with a positively charged Arg or negatively charged Glu, resulted in the predicted increase or decrease in reduction potential at all pH values. Similarly, substitution of Met121 with a more hydrophobic Leu raised the reduction potential. The increased solubility, stability, and tune-ability of this organometalloprotein make it an ideal choice for carrying out a number of biological reactions, such as long-range electron transfer or sensing. As an example of such applications, stoichiometric oxidation of ferrocytochrome c by the blue ferrocenium azurin was demonstrated.     

Two-dimensional electrophoresis reveals differential protein expression in high- and low-secreting variants of the rat basophilic leukaemia cell line

The aim of this investigation was the identification of cellular proteins that confer a high secretory phenotype on subclones of the rat basophilic leukaemia (RBL) cell line as a model of mast cell regulated degranulation. Following protein separation by two-dimensional (2-D) electrophoresis and silver staining, more than 2000 polypeptide "spots" were resolved reproducibly. Higher sample loads and Coomassie blue staining facilitated the identification by delayed extraction-matrix-assisted laser desorption/ionization (DE-MALDI) mass spectrometry of several polypeptides that were differentially expressed in the high- and low-secreting clones. Several proteins were identified whose expression could contribute to the difference in secretory phenotype. Furthermore, silver-stained 2-D gel patterns suggested differential expression of proteins in the 20-25 kDa and the pI 4.5-7.5 range, characteristic of small guanosine 5'-triphosphate (GTP)-binding proteins. By a combination of "GTP overlay" and immunoblotting, we were able to demonstrate differential expression of small GTP binding-proteins, including Rab3 proteins, in high-and low-secreting clones. The sensitivity of this complementary approach facilitated the detection of some GTP binding and Rab3 proteins, whose expression was not evident in silver-stained 2-D gels.