Determination of interproton distances in peptides by two-dimensional nuclear Overhauser effect spectroscopy (NOESY). Conformation of a cyclic analog of substance P in a solution

Quantitative method is developed for evaluation interproton distances in peptides in solution. The method is based on the measurement of the relative intensities of the cross-peaks in the pure-phase absorption NOESY spectra. The ratios of the cross-peak intensities IN alpha/I alpha N and INN/I alpha N enable to determine the corresponding interproton distances dN alpha, d alpha N and dNN for several amino acid residues. These distances can be used to estimate other distances with cross-peaks in NOESY spectra. As example, the interproton distances are determined in a cyclic hexapeptide, namely cyclic analogue of substance P: cyclo [H-Glu-Phe-Phe-Gly-Leu-Met-NH(CH2)3-NH-]. The spatial structure of the molecule in dimethylsulphoxide solution is established.     

A new species of Bradyidius (Copepoda, Calanoida) from the Gulf of Carpentaria, Australia

A new species of copepod belonging to the family Aetideidae,Bradyidius slyliformis sp. nov. is described and figured. Therelationship of this species to the other 12 members of thegenus is discussed. ¹Present address: 121 Jalan Athinahapan Dua, Taman Tun Dr Ismail,60000 Kuala Lumpur, Malaysia     

Effect of high concentration of nutrients onBacillus thuringiensis cultures

Summary Bacterial insecticide production using a strain ofBacillus thuringiensis var. kurstaki was studied in batch culture considering the influence of increasing concentration of components of a glucose — yeast extract — mineral salts medium.It was found that spore counts were increased from 1.08×10¹² spores. 1^–1 to 7.36×10¹² spores. 1^–1 and toxin level from 1.05 mg.ml^–1 to 6.85 mg.ml^–1, when the concentration of glucose was increased from 8 to 56 (g 1^–1), with the corresponding increase in the rest of medium components. Higher concentration of nutrients inhibit either spore count or toxin production.Preliminary experiments of fed-batch cultures which allows the use of high amounts of nutrients were also carried out. In this case spore counts of 1.2×10¹³ spores.1^–1 were achieved.     

Characterization of a light-dependent glutamate synthase activity in Chlamydomonas reinhardtii

Photosynthetically active vesicles prepared from Chlamydomonas reinhardtii retained a light-dependent glutamate synthase activity which was highly specific for 2-oxoglutarate (K_m=2.1 mM) and L-glutamine (K_m=0.9 mM) as amido group acceptor and donor respectively. This activity was inhibited by azaserine, p-hydroxymercuribenzoate and 3-(p-chlorophenyl)-1,1-dimethyl urea.Light-dependent synthesis of glutamate was also obtained by coupling Chlamydomonas photosynthetic particles to purified ferredoxin-glutamate synthase, using ascorbate and 2,6-dichlorophenol-indophenol as electron donor. This system was also specific for 2-oxoglutarate (K_m=1 mM) and L-glutamine (K_m=0.8 mM) as substrates, and was stimulated by dithioerythritol. Azaserine and p-hydroxymercuribenzoate, but not 3-(p-chlorophenyl)-1,1-dimethyl urea, inhibited the reconstituted activity; high concentrations of 2-oxoglutarate were inhibitory.Abbreviations A Absorbance - CCP p-Trichlorometoxi-carbonylcyanide-phenylhydrazone - Chl Chlorophyll - CMU 3-(p-Chlorophenyl)-1,1-dimethyl urea - DPIP 2,6-Dichlorophenol-indophenol - DTE Dithioerythritol - MSX L-Methionine, D-L, sulfoximine - MV Methyl viologen     

Pheophytin-mediated energy storage of photosystem II particles detected by photoacoustic spectroscopy

The photoacoustic (PA) characteristics (energy storage and heat dissipation) of photosystem II (PSII) core-enriched particles from barley were studied (i) in conditions where there was electron flow, i.e., in the presence of a combination of the electron acceptor K₃ Fe (CN)₆, referred to as FeCN, and the electron donor diphenylcarbazide (DPC), and (ii) in conditions where electron flow was suppressed, i.e., in the absence of FeCN and DPC. The experimental data show that a decrease of heat dissipation with a minimum at 540 nm can be interpreted as energy storage resulting from the presence of pheophytin (Pheo) in the PSII particles. On account of the capability of the PA method to measure the energy absorbed by the chromophores which is converted to heat, it is suggested that the PA detection of Pheo present in the PSII complex will permit to clarify the function of processes involving non-radiative relaxation of excited states in P680-Pheo-Q_A interactions.Abbreviations -Car -Carotene - Chl Chlorophyll - DPC Diphenylcarbazide - EPR Electron Paramagnetic Resonance - FeCN potassium ferricyanide - HEPES N-2-hydroxyethylenepiperazine-N-2-ethanesulfonate - P680 reaction center of PSII - PA Photoacoustic - Pheo pheophytin - PSI photosystem I - PSII photosystem II - Q_A primary electron acceptor of PSII     

The biogeochemical cycle of the adsorbed template

Experimental results are presented for the verification of the first adsorption step of the ‘adsorbed template’ biogeochemical cycle, a simple model for a primitive prebiotic replication system. The adsorption of Poly-C, Poly-U, Poly-A, Poly-G, and 5′-AMP, 5′-GMP, 5′-CMP and 5′-UMP onto gypsum was studied. It was found that under the conditions of the experiment, the polymers have a very high affinity for the mineral surface, while the monomers adsorb much less efficiently.     

Purification of bovine liver histidyl-tRNA synthetase, the Jo-1 antigen of polymyositis: size of the whole enzyme and its characteristic proteolytic fragments

We have recently reported the marked increase in frequency which can be achieved in the detection of the anti-Jo-1 antibody of polymyositis in serum samples by replacing commercial mixtures of cytoplasmic and nuclear antigens with the purified antigen, histidyl-tRNA synthetase. The present paper describes a method for purifying this antigen and an investigation of its size. Molecular masses previously reported for the enzyme have varied from 85-154 kDa and subunit molecular masses varying from 40-77 kDa have been observed. Several of these fragments are of sizes similar to those of a number of other autoantigens commonly observed in connective tissue diseases. Since the clinical identification of these autoantigens often relies exclusively on size determination by Western blotting, we have characterized the commonly occurring fragments of histidyl-tRNA synthetase lest they confuse such identification. It is concluded that histidyl-tRNA synthetase, like many other aminoacyl-tRNA synthetases, is subject to severe proteolysis during extraction procedures. Several characteristic fragments (Mr = 80, 75, 61, 55, 50 and 45 kDa) result, a finding that provides a satisfactory explanation of the various values previously reported. The intact bovine enzyme is a dimer of molecular mass close to 160 kDa.     

Investigations on circumferential microfilament bundles in rat retinal pigment epithelium

The presence of contractile proteins in normal rat retinal pigment epithelium has been studied using fluorescence and electron microscopy. Investigations using the F-actin binding toxin, phallacidin, coupled to the fluorochrome nitrobenzoxadiazole, revealed a band of fluorescence at or near the cell membrane. Immunofluorescent observations with anti-myosin and anti-alpha-actinin antisera gave similar results. Electron microscopy employing glutaraldehyde-8% tannic acid fixation revealed the presence of a circumferential microfilament band beneath the pigment epithelial apical surface that is closely associated with the plasma membrane and junctional complexes. Freeze-fracture studies confirmed the relationship of this band to the junctional complexes. The microfilament band measures approximately 0.5 micron +/- 0.2 micron in width and is composed of numerous 6 to 7 nm filaments. Some microtubules are seen in regions around the band, but no organelles appear to be associated with this structure. In en face sections through the zonula adherens, the circumferential microfilament band is associated with 30-nm electron-dense particles that are bound to the internal side of the membrane. Morphological evidence suggests that these may serve in anchoring the band to the membrane and assist in aligning the microfilament bands of adjoining cells. In the subapical cytoplasm, a microfilament bundle network was detected that interfaced with the circumferential microfilament band. In some cases, pigment epithelium was incubated in media-199 containing 25 to 50 ng/ml phallacidin prior to fixation. Circumferential microfilament bands of tissues treated in this manner exhibited a striated appearance.     

Genetic study on indole-3-acetic acid production by ectomycorrhizal Hebeloma species: inter- and intraspecific variability in homo- and dikaryotic mycelia

Summary Interspecific variability of indole-3-acetic acid (IAA)-synthesizing activity was examined within 12 wild strains of different Hebeloma species. Interstrain variability was studied within 11 wild strains of Hebeloma cylindrosporum (Romagnési) and intrastrain variability was considered by using 20 homokaryotic and 50 controlled dikaryotic mycelia belonging to the progeny of one laboratory fruiting strain of this species.The range of variation of IAA-synthesizing activity was of the same order of magnitude within the four groups considered. No correlation was detected between, on one hand, the IAA-synthesizing activity of the mycelia and, on the other hand, their taxonomic position, their geographic origin, or their host plant.Within the progeny of one H. cylindrosporum fruiting strain, 15 of the 50 controlled dikaryons presented an activity higher than that of the original dikaryon. The variation among dikaryons could not be strictly related to the variation in parental homokaryons, indicating that genetic control of this activity probably involves a nonadditive component. Significant additive and nonadditive components of the genetic variation were detected, each of them representing about 50% of the total variation. The nonadditive heritable component could not be explained by a model involving only dominance.