Metabolism of 2,4-dichlorophenoxyacetic acid in cell suspension cultures of soybean (Glycine max L.) and wheat (Triticum aestivum L.)

Cell suspension cultures of soybean (Glycine max L.) and wheat (Triticum aestivum L.) incorporated 2,4-dichlorophenoxyacetic acid (2,4-D) into a metabolite fraction which was insoluble in ethanol, water, and hot sodium dodecylsulphate. Further treatment with hot dimethylformamide solubilized a material which by the following criteria appeared to consist of 2,4-D derivatives covalently bound to lignin: i) co-chromatography of radioactivity and of UV-absorbing material upon gel permeation chromatography; ii) spectral similarity with authentic lignins (IR- and UV-spectra, phloroglucinol reaction), 2,4-D appeared to be incorporated as the intact molecule, as shown by comparison of ring- and sidechain-labeled 2,4-D and by detection of monohydroxylated and intact 2,4-D as the major radioactive products of acid hydrolysis. The same compounds were released from the metabolite material which could not be solubilized in dimethylformamide. The incorporation of xenobiotics or their metabolites into lignin, followed by deposition in the cell wall, is suggested as a general pathway for local excretion and detoxification by plant cells.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - 4-OH-2,5-D 4-hydroxy-2,5-dichlorophenoxyacetic acid - SDS sodium dodecylsulphate - DMF dimethylformamide     

Über den submikroskopischen Bau des Grundhäutchens der Hirnkapillaren

Zusammenfassung Auf Grund der polarisationsoptischen Analyse besitzt das homogene Kapillar-Grundhäutchen eine geordnete Feinstruktur. Es ist ein Eiweiß-Lipoidsystem, in dem die Proteinkomponente als Träger- oder Gerüstsubstanz mengenmäßig überwiegt. Die Proteine zeigen teilweise eine fibrilläre Struktur mit geringer mizellarer Orientierung. In der Fläche der Membran haben die Fibrillen keine bevorzugte Verlaufsrichtung, sie liegen statistisch ungeordnet. Zu der Membranoberfläche verlaufen sie jedoch annähernd parallel, so daß eine folienartige Textur resultiert. In dieses Proteingerüst sind Lipoidmolekeln in radiärer Richtung orientiert eingelagert. Nach einer Näherungsrechnung dürfte es sich um eine bimolekulare Lamelle handeln, wahrscheinlich ist, daß die Lipoide inselförmig in das Proteingerüst eingesprengt sind. Die gefundene Struktur wird in Beziehung zu den Befunden neuerer Arbeiten über die Blut-Hirnschranke und die Blut-Liquorschranke gesetzt. Hierbei wird die Bedeutung der Membranlipoide für die Regulierung des Durchtritts lipoidlöslicher Stoffe, diejenige der Proteinlamellen für die Regulierung der Durchlässigkeit für Wasser und wasserlösliche Stoffe erörtert. Die Endothelmembran, d. h. das Grundhäutchen der Kapillare wird als das morphologische Substrat der Schrankenfunktion angesehen.Die Untersuchung wurde mit dankenswerter Unterstützung der Deutschen Forschungsgemeinschaft ausgeführt.     

She2p is a novel RNA binding protein with a basic helical hairpin motif

Selective transport of mRNAs in ribonucleoprotein particles (mRNP) ensures asymmetric distribution of information within and among eukaryotic cells. Actin-dependent transport of ASH1 mRNA in yeast represents one of the best-characterized examples of mRNP translocation. Formation of the ASH1 mRNP requires recognition of zip code elements by the RNA binding protein She2p. We determined the X-ray structure of She2p at 1.95 A resolution. She2p is a member of a previously unknown class of nucleic acid binding proteins, composed of a single globular domain with a five alpha helix bundle that forms a symmetric homodimer. After demonstrating potent, dimer-dependent RNA binding in vitro, we mapped the RNA binding surface of She2p to a basic helical hairpin in vitro and in vivo and present a mechanism for mRNA-dependent initiation of ASH1 mRNP complex assembly.     

The ubiquinone-binding site in NADH:ubiquinone oxidoreductase from Escherichia coli

An azido-ubiquinone derivative, 3-azido-2-methyl-5-methoxy[3H]-6-decyl-1,4-benzoquinone ([3H]azido-Q), was used to study the ubiquinone/protein interaction and to identify the ubiquinone-binding site in Escherichia coli NADH:ubiquinone oxidoreductase (complex I). The purified complex I showed no loss of activity after incubation with a 20-fold molar excess of [3H]azido-Q in the dark. Illumination of the incubated sample with long wavelength UV light for 10 min at 0 degrees C caused a 40% decrease of NADH:ubiquinone oxidoreductase activity. SDS-PAGE of the complex labeled with [3H]azido-Q followed by analysis of the radioactivity distribution among the subunits revealed that subunit NuoM was heavily labeled, suggesting that this protein houses the Q-binding site. When the [3H]azido-Q-labeled NuoM was purified from the labeled reductase by means of preparative SDS-PAGE, a 3-azido-2-methyl-5-methoxy-6-decyl-1,4-benzoquinone-linked peptide, with a retention time of 41.4 min, was obtained by high performance liquid chromatography of the protease K digest of the labeled subunit. This peptide had a partial NH2-terminal amino acid sequence of NH2-VMLIAILALV-, which corresponds to amino acid residues 184-193 of NuoM. The secondary structure prediction of NuoM using the Toppred hydropathy analysis showed that the Q-binding peptide overlaps with a proposed Q-binding motif located in the middle of the transmembrane helix 5 toward the cytoplasmic side of the membrane. Using the PHDhtm hydropathy plot, the labeled peptide is located in the transmembrane helix 4 toward the periplasmic side of the membrane.