Zur Biosynthese der C9–10-Einheit der Indolalkaloide

Summary After administration of various C₂-compounds as well as leucine-2-¹⁴C toCatharanthus roseus shoots and glycine-2-¹⁴C toStrychnos nux vomica plants no specific incorporation into the non-tryptophan C_9–10 moiety of indole alkaloids was observed. The results indicate that glycine-2-¹⁴C is transformed into serine and is incorporated via tryptophan into strychnine.     

Taxuspine D,a new taxane diterpene fromTaxus cuspidata with potent inhibitory activity against Ca2+-induced depolymerization of microtubules

A new taxane diterpenoid, taxuspine D (1), possessing an enolacetate moiety, has been isolated from stems of the Japanese yewTaxus cuspidata Sieb. et Zucc., and the structure elucidated on the basis of spectroscopic data. Taxuspine D (1) markedly inhibited Ca²⁺-induced depolymerization of microtubules.     

Synthesis of L-[3-2H,18O]glycerol and its incorporation into the 4-methyl-5-hydroxyethyl thiazole moiety of thiamine byEscherichia coli

Summary L-[3-²H,¹⁸O]glycerol was prepared and fed toEscherichia coli in order to determine the origin of the oxygen atom in the biosynthesized thiazole moiety of thiamine. Measurement by GC-MS of the isotope incorporation into the thiazole from this substrate confirmed that the 2 hydrogens and the oxygen on the C-3 carbon of glycerol are incorporated directly into the thiazole.     

Zur Biosynthese des Plumierids

Summary It has been shown that the leaves ofPl. acutifolia incorporate acetate-1-¹⁴C and mevalonate-2-¹⁴C specifically for the formation of the aglycone moiety of plumieride.

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Wir danken dem Schweizerischen Nationalfonds sehr für die Unterstützung der Arbeit, Herrn Dr.J. Würsch (F. Hoffmann-La Roche & Cie., Basel) für markiertes Mevalonsäure-lakton und Herrn Prof.D. Arigoni (Zürich) für interessante Diskussionen.     

A novel cannabinoid containing a 1,8-cineol moiety

Summary The synthesis of a novel cannabinoid containing a 1,8-cineol moiety (1) is described. It is much less biologically active than ¹-tetrahydrocannabinol.Acknowledgment. This work was supported by NIDA (grant No. DA-00574-01).     

The structure and function of platelet-activating factor acetylhydrolases

Platelet-activating factor acetylhydrolases (PAF-AHs, EC 3.1.1.47) constitute a unique and biologically important family of phospholipase A₂s. They are related to neither the well-characterized secretory nor cytosolic PLA₂s, and unlike them do not require Ca²⁺ for catalytic activity. The distinguishing property of PAF-AHs is their unique substrate specificity they act on the phospholipid platelet-activating factor (PAF), and in some cases on proinflammatory polar phospholipids, from which they remove a short acyl moiety – acetyl in the case of PAF – located at the sn-2 position. Because PAF is found both in the plasma and in the cytosol of many tissues, PAF-acetylhydrolases are equally widely distributed in an animal organism. Recent crystallographic studies shed new light on the complex structure-function relationships in PAF-AHs. Received 15 September 1997; received after revision 23 February 1998; accepted 25 February 1998     

DDT: The degradation of ring-labeled14C-DDT to14CO2 in the rat

Summary Ring fission ofp, p-DDT was studied in the rat following a single oral dose of 0.74 mg/kg (1.04 Ci) of uniformly ring-labeled¹⁴C-DDT. Expired air was passed through a solution of ethanolamine-ethylene glycol monomethyl ether (12) to trap¹⁴CO₂. A total of 1.6% of the radioactivity administered was recovered in the expired air collected continually for 10 days, indicating that while degradation of the phenyl moiety is not a major route pfp, p-DDT metabolism in the rat, it is equal to the urinary excretion. Nevertheless, these results represent the most radical change accomplished in vivo of a residual insecticide yet reported in mammals.Acknowledgment. We acknowledge the secretarial work of Ms.Elaine Smolko. This study was supported by NIH fellowship No. 1 F22 ES01723-01 and No. HL16264.     

On the synthesis of serine and homoserine samples asymmetrically labelled with tritium and deuterium in the hydroxymethylene group

Summary (1 R) [1-³H,²H₁] 3-Phenylpropanol, the key intermediate in the synthesis of (4 R) [4-³H,²H₁] D, L-homoserine and of the (4 S)-isomer, is obtained from (1 S) [1-²H₁] 3-phenylpropanol and (1 RS) [1-³H] ethanol upon incubation with yeast alcohol dehydrogenase and NAD⁺; under similar conditions 2-phenylethanol undergoes very small exchange with [1-²H₂] ethanol.     

Elektronenspinresonanzmessungen zur Untersuchung von Kinetik und Mechanismus von Cu2+-katalysierten Reaktionen

Summary Cu²⁺-complexes with different monodentate ligands PYR, e.g. pyridine, 2,4,6-collidine and imidazole, catalyse the oxidation ofo-phenylenediamine (H₂B) to 3,5-dihydro-2-amino-3-iminophenazine (PHEN) by O₂. Investigation of the electron paramagnetic resonance during reaction gives interesting details on the function of Cu²⁺ as a catalyser. The formation of mixed complexes (H₂B)Cu²⁺(PYR) and its influence on the reaction rated[PHEN]/dt is demonstrated. In the ratedetermining reaction, Cu²⁺ is reduced to Cu⁺, which is reoxidized by O₂. During reaction the ratio [Cu²⁺]/[Cu⁺] is determined by means of e.p.r. measurements.     

In pre-sterol carrier protein 2 (SCP2) in solution the leader peptide 1 – 20 is flexibly disordered, and residues 21 – 143 adopt the same globular fold as in mature SCP2

The preform of the rabbit sterol carrier protein 2 (pre-rSCP2) was cloned, the uniformly ¹⁵N-labelled protein expressed in Escherichia coli and studied by three-dimensional ¹⁵N-resolved nuclear magnetic resonance spectroscopy. In spite of its low solubility in aqueous solution of only ∼0.3 mM, sequential ¹⁵N and ¹H backbone resonance assignments were obtained for 105 out of the 143 residues. From comparison of the sequential and medium-range nuclear Overhauser effects (NOEs) in the two proteins, all regular secondary structures previously determined in mature human SCP2 (hSCP2) [Szyperski et al. (1993) FEBS Lett. 335: 18–26] were also identified in pre-rSCP2. Near-identity of the backbone ¹⁵N and ¹H chemical shifts and 1 : 1 correspondence of 24 long-range NOEs to backbone amide groups in the two proteins show that the residues 21 – 143 adopt the same globular fold in pre-rSCP2 and mature hSCP2. The N-terminal 20-residue leader peptide of pre-rSCP2 is flexibly disordered in solution and does not observably affect the conformation of the polypeptide segment 21 – 143. Received 11 May 1998; accepted 15 May 1998     

Zur Frage der Beziehungen zwischen enddiastolischem Ventrikeldruck unddp/dt bzw.d 2 p/dt 2 der Ventrikeldruckkurve des Herzens

Summary Correlations between left ventricular enddiastolic pressure of the heart and first and second derivation of left ventricular pressure curve (dp/dt, d ² p/dt ² respectively) were examined at various hemodynamic conditions. The results showed only a particular dependence ofdp/dt andd ² p/dt ² from enddiastolic pressure. Therefore, it is concluded, that enddiastolic pressure is not the most important factor, which influences the dynamics of ventricular contraction.     

Zur Stereochemie der Propandioldehydrase-Reaktion

Summary Propanedioldehydrase is shown to convert both (+)-(S)-2-²H-propanediol,3, and (–)-(R)-1-²H₂-propanediol,5, to specimens of deuterated propion-aldehyde, for which the (S)-configuration has been established. Thus, in the propanedioldehydrase reaction migration of hydrogen atoms from C–1 to C–2 always occurs with inversion of configuration.     

The superoxide-generating NADPH oxidase: structural aspects and activation mechanism

Flavocytochrome b ₅₅₈ is the catalytic core of the respiratory-burst oxidase, an enzyme complex that catalyzes the NADPH-dependent reduction of O₂ into the superoxide anion O₂ ^- in phagocytic cells. Flavocytochrome b ₅₅₈ is anchored in the plasma membrane. It is a heterodimer that consists of a large glycoprotein gp91phox (phox for phagocyte oxidase) (β subunit) and a small protein p22phox (α subunit). The other components of the respiratory-burst oxidase are water-soluble proteins of cytosolic origin, namely p67phox, p47phox, p40phox and Rac. Upon cell stimulation, they assemble with the membrane-bound flavocytochrome b ₅₅₈ which becomes activated and generates O₂ ^-. A defect in any of the genes encoding gp91phox, p22phox, p67phox or p47phox results in chronic granulomatous disease, a genetic disorder characterized by severe and recurrent infections, illustrating the role of O₂ ^- and the derived metabolites H₂O₂ and HOCl in host defense against invading microorganisms. The electron carriers, FAD and hemes b, and the binding site for NADPH are confined to the gp91phox subunit of flavocytochrome b ₅₅₈ . The p22phox subunit serves as a docking site for the cytosolic phox proteins. This review provides an overview of current knowledge on the structural organization of the O₂ ^--generating flavocytochrome b ₅₅₈ , its kinetics, its mechanism of activation and the regulation of its biosynthesis. Homologues of gp91phox, called Nox and Duox, are present in a large variety of non-phagocytic cells. They exhibit modest O₂ ^--generating oxidase activity, and some act as proton channels. Their role in various aspects of signal transduction is currently under investigation and is briefly discussed. Received 28 May 2002; received after revision 20 June 2002; accepted 24 June 2002     

Direct inhibition of phospholipid scrambling activity in erythrocytes by potassium ions

The exposure of phosphatidylserine (PS) at the cell surface plays a critical role in blood coagulation and serves as a macrophage recognition moiety for the engulfment of apoptotic cells. Previous observations have shown that a high extracellular [K⁺] and selective K⁺ channel blockers inhibit PS exposure in platelets and erythrocytes. Here we show that the rate of PS exposure in erythrocytes decreases by ~50% when the intracellular [K⁺] increases from 0 to physiological concentrations. Using resealed erythrocyte membranes, we further show that lipid scrambling is inducible by raising the intracellular [Ca²⁺] and that K⁺ ions have a direct inhibitory effect on this process. Lipid scrambling in resealed ghosts occurs in the absence of cell shrinkage and microvesicle formation, processes that are generally attributed to Ca²⁺-induced lipid scrambling in intact erythrocytes. Thus, opening of Ca²⁺-sensitive K⁺ channels causes loss of intracellular K⁺ that results in reduced intrinsic inhibitory effect of these ions on scramblase activity. Received 11 September 2008; received after revision 17 October 2008; accepted 27 October 2008     

Developmental changes of aldehyde dehydrogenase isozymes in human livers: Mitochondrial ALDH2 isozyme is expressed in fetal livers

Summary Previous reports suggested that the major cytosolic aldehyde dehydrogenase (ALDH₁) was present in fetal and infant livers, but the major mitochondrial isozyme (ALDH₂) was absent or severely diminished. Re-examination by means of starch gel electrophoresis followed by enzyme activity staining, and by means of dot blot immuno-hybridization of liver samples with known genotypes of theALDH ₂ locus, indicated that bothALDH ₁ andALDH ₂ genes are expressed in fetal and infant livers. In addition, ALDH₄ isozyme was also observed. The results imply that a fetus with the usual homozygousALDH ₂ ¹ /ALDH ₂ ¹ genotype, but not one with the atypicalALDH ₂ ¹ /ALDH ₂ ² orALDH ₂ ² /ALDH ₂ ² , is capable of detoxifying acetaldehyde transferred from the mother.     

Blockade by zinc ions of K+-induced contraction and calcium in guinea pigTaenia coli

Preincubation with 0.3 mM Zn²⁺ markedly inhibited both the tonic response and Ca²⁺ binding at low affinity sites induced by K⁺ (60 mM), with smaller effects on the phasic response and the high affinity Ca²⁺ sites, inTaenia coli. However, when the muscle was kept in Zn²⁺-containing medium following the first stimulation with the K⁺, the phasic response and the high affinity Ca²⁺ sites were more severely inhibited during the second stimulation with K⁺. This probably indicates that Zn²⁺ reduced the tonic tension response to K⁺ mainly by inhibiting Ca²⁺ influx at the cell membranes ofTaenia coli. However, when Zn²⁺ is continuously present, Ca²⁺ is not supplied at the storage sites and is not available for the phasic response to a second stimulation with K⁺.     

Gitterkonstanten und Raumgruppe von Tetrammincuprisulfat

Summary The space-group of Cu(NH₃)₄SO₄·H₂O isD _2h ¹⁶ orD _2h ¹³ (D _2h ⁵ , D _2h ¹ ) with a=7,07, b=12,12, c=10,66 Å ± 1%.     

Observation of pentacoordinated phosphorus intermediate in the reactions of nitrones with phosphonate anions

Summary The occurrence of a high-field signal in the³¹P FT NMR-spectrum of the reaction mixture of nitrones (1 or2) and 2-cyanomethyl-4,5-dimethyl-2-oxo-1,3,2-dioxyphospholane (7), is interpreted in terms of a pentacoordinated phosphorus intermediate.     

Arachidonic acid release by ionomycin and phorbol ester is similar in C127 epithelial cells expressing wild-type or mutated (ΔF508) cystic fibrosis transmembrane conductance regulator

The Ca²⁺ ionophore ionomycin induced cytosolic [Ca²⁺]_i elevation as well as strong activation of Cl⁻ efflux in mouse mammary epithelial cell lines expressing wild-type or mutated (deletion of phenylalaline 508) cystic fibrosis transmembrane conductance regulator (CFTR) or vector. Ionomycin-induced Cl⁻ efflux was abolished by the intracellular Ca²⁺ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid, whereas both activators and inhibitors of phospholipase A₂ had no effect, indicating the involvement of Ca²⁺-dependent Cl^- channels. Stimulation of arachidonic acid release by ionomycin and phorbol ester was not significantly different between wild-type or mutated cell lines, whereas vector-transfected cells exhibited a significant higher release, which was shown to be due to larger amount of immunoreactive cytosolic phospholipase A₂. These results indicate that phospholipase A₂ activity of C127 cells was not influenced by the presence of wild-type or mutated CFTR. Received 27 April 1999; received after revision 11 June 1999; accepted 23 July 1999