Separation of friend erythroleukaemic cell histones and high-mobility-group proteins by reversed-phase high-performance liquid chromatography

A procedure for the rapid separation of histones and high-mobility-group (HMG) proteins from Friend erythroleukaemic cells (line F4N) by reversed-phase high-performance liquid chromatography is reported. By using a Nucleosil 300-5 C₄, column and a multistep water—acetonitrile gradient containing 0.1% trifluoroacetic acid, the HMG-1 and HMG-2 proteins, several his subtractions including H1⁰, H4, H2B, two H2A variants and two H3 subtractions were separated. Under changed conditions, by applying a varied acetonitrile gradient system, even two H2B variants were fractionated. The methods described seem to be a real alternative to the time-consuming polyacrylamide gel electrophoresis.     

Molekül- und Kristallstruktur des 1,4-Bis[tris(tetrahydrofuran)lithium]-octaphenyltetrasilans

Molecular and Crystal Structure of 1,4-Bis[tris(tetrahydrofuran)lithium]-octaphenyltetrasilane 1,4-Dilithium-octaphenyltetrasilane prepared from octaphenyl-cyclo-tetrasilane and lithium in tetrahydrofuran (THF) [4], can be isolated from tetrahydrofuran/n-pentane as an adduct with six molecules of tetrahydrofuran per formula unit. The orange-red compound crystallizes in the triclinic space group P1 {a = 1159.6(3); b = 1268.4(2); c = 1367.8(3) pm; α = 92,23(2)° β = 113.79(2)° γ = 111.62(2)° at ?5 ± 3°C; Z = 1}. An x-ray structure determination (R_w = 0.046) shows the existence of a centrosymmetric molecule with an extended planar Li? Si₄? Li unit; either lithium atom is bound to silicon and to the oxygen atoms of three molecules of tetrahydrofuran. Characteristic bond lengths and angles are: Li? Si 271; Si? Si 241 and 243; Si? C 190 to 192 pm; Li? Si? Si 126°; Si? Si? Si 127°. ²⁹Si and ⁷Li n.m.r. measurements at low temperatures indicate the presence of three different adducts.     

Electrochemical oxidation of alkyl-substituted allenes in methanol

Mono-, di- and tri-alkyl-substituted allenes were potentiostatically oxidized in methanol at graphite and Pt anodes. At the former electrode, α-methoxylated ketones (due to 4F/mole electricity consumption) and esters (6F/mole) were the major products. At a Pt anode, intermediate products such as vinyl-ether derivatives (2F/mole) were characterised too. Unlike the anodic oxidation of alkenes and alkynes previously reproted in the literature, dimerisation is not a typical process in the allenes' oxidation, since of all the products obtained only a sole dimer has been observed. The mechanism for the formation of most products is discussed.     

Herstellung und Eigenschaften von geformtem Aluminiumoxid. I. Einfluß der Fällbedingungen des Böhmithydrogels auf die Porenstruktur des geformten Aluminiumoxids

Preparation and Properties of Formed Aluminium Oxide. I. Influence of the Precipitation Conditions of the Boehmite Hydrogel on the Pore Structure of Formed Aluminium Oxide A report is given on the influence of the precipitation conditions of boehmite (pH value, temperature, concentration and residence time in the precipitation suspension) on the cavity structure of aluminium oxide spheres, made by coagulation of boehmite hydrosol in ammonia liquor and subsequent thermal treatment at 110 and 600°C. The boehmite hydrogel was obtained at continuous precipitation conditions by neutralisation of sodium aluminate solution with nitric acid. It is shown that the difference in the pore structure of the formed aluminium oxide obtained by varying the precipitation conditions were caused by the special morphological features of the boehmite crystallization in the precipitation process.     

Electrode processes in the oxidation of tetrahydroisoquinoline derivatives

The electrochemical oxidation of the alkaloid laudanosine (Ia) to O-methylflavinantine (II) has been studied in acetonitrile solvent. Using cyclic voltammetry, rotating disc voltammetry and preparative electrolyses on several alkaloids, simple aliphatic amines and aromatic compounds, some aspects of the mechanism of this coupling reaction are elucidated. The first anodic wave for laudanosine at platinum has E_p=0.55 V vs. Ag/Ag⁺. The electrode rapidly becomes partially passivated at potentials above 0.5 V. This is due to a film which “dissolves” below 0.5 V, at a rate independent of the potential. It is shown that the reaction (Ia)→(II) proceeds at 0.5 V by initial oxidation of the amine moiety. If acids such as sodium bicarbonate are added to the anolyte the amine is protonated causing the first wave to disappear. Oxidation at 1.1 V under these acidic conditions produces the same product, but more rapidly and in significantly higher yield because electrode filming and side reactions resulting from the amine oxidation are abrogated.     

Biosynthesis of 4-methylproline in cyanobacteria: cloning of nosE and nosF genes and biochemical characterization of the encoded dehydrogenase and reductase activities

The biosynthesis of the unusual amino acid 4-methylproline in the Nostoc genus of cyanobacteria was investigated on the genetic and enzymatic level. Two genes involved in the biosynthesis were cloned and the corresponding enzymes, a zinc-dependent long-chain dehydrogenase and a Delta(1)-pyrroline-5-carboxylic acid (P5C) reductase homologue, were overexpressed in Escherichia coli and biochemically characterized. Putative substrates were synthesized to test enzyme substrate specificities, and deuterium labeling studies were carried out to reveal the stereospecificities of the enzymatic reactions with respect to the substrates as well as to the coenzymes.     

Acyl- und Alkylidenphosphane. XXVII [I.]. Molekül- und Kristallstruktur des Methyl[(N-phenyl,N-tri-methylsilyl)thiocarbamoyl]trimethylsilylphosphans

Acyl- and Alkylidenephosphines. XXVII. Molecular and Crystal Structure of Methyl-[(N-phenyl, N-trimethylsilyl)thiocarbamoyl]trimethylsilylphosphine . Methyl[(N-phenyl, N-trimethylsilyl)thiocarbamoyl]trimethylsilylphosphine 1a formed via an addition of methylbis(trimethylsilyl)phosphine to phenyl isothiocyanate [1], crystallizes in the monoclinic centrosymmetric space group P2₁/n with following dimensions of the unit cell determined at a temperature of measurement of ?80±3°C: a=1041.2(4);b=1706.9(12);c=1001.1(6)pm; β=106.41(4)°; Z = 4. An X-ray structure determination (R_w = 0.039) confirms the constitution of the compound as already derived from its nmr spectra. One trimethylsilyl group is bound to the phosphorus atom, whereas the other is connected with the sp²-hybridized nitrogen atom. Characteristic rounded bond lenghts and angles are: P? Si 231, P? CH₃ 184, P? C(S) 187, C?S 167, N? C(S) 137, and N? Si 181 pm as well as P? C? S 122°, P? C? N 117°, and S? C? N 121°.     

Comparative studies of MCs+-SIMS and e–-beam SNMS for quantitative analysis of bulk materials and layered structures

For the quantification of heterostructure depth profiles the knowledge of relative sensitivity factors (RSF) and the influence of matrix effects on the measured profiles is necessary. Matrix dependencies of the measured ion intensities have been investigated for sputtered neutral mass spectrometry (SNMS) and MCs(+)-SIMS. The use of Cs as primary ions for SNMS is advantageous compared to Ar because the depth resolution is improved without changing RSFs determined under Ar bombardment. No significant amount of molecules has been found in the SNMS spectra under Cs bombardment. Using MCs(+)-SIMS the RSFs are matrix dependent. An improvement of depth resolution can be achieved by biasing the sample against the primary ion beam for SNMS due to a reduction of the net energy of the primary ions and a resulting more gracing impact angle.     

Quantification of free and total salicylic acid in plants by solid-phase extraction and isocratic high-performance anion-exchange chromatography

Salicylic acid (SA) is an important signaling compound in plants and is involved in various defense responses. Here we report a new method for quantification of free and total soluble SA in Arabidopsis thaliana with 5-fluorosalicylic acid (5-FSA) as internal standard. The SA was isolated from leaf extracts by solid-phase extraction with phenyl-phase cartridges and selectively eluted as the cationic iron(III)-complex. Recoveries of SA and 5-FSA were equal and exceeded 90%. Free SA was subsequently released from the iron(III)-complex by addition of 2,2-bipyridyl and high-performance anion-exchange chromatography was performed on an NH₂ column. The SA appeared as last peak with a retention time of 15 min, baseline-separated from other substances. On-line detection was performed fluorimetrically for both SA and 5-FSA at an excitation wavelength of 300 nm and an emission wavelength of 410 nm, because both substances give similar fluorescence spectra. The detection limit for SA was 5 ng g^–1 FW for a sample size of 100 mg. Thus the main advantages of the method are highly selective sample preparation, increased sensitivity, reduced analysis time compared with reversed-phase HPLC, and use of a novel internal standard detectable under the same conditions as SA. The techniques described are applicable to other plant materials.