Conformational Changes of αs-Casein by Heating

Conformational changes of α_s-casein by heating were investigated by measuring ultraviolet difference spectra. The ultraviolet difference spectra at elevated temperature against 5.5°C were measured in various ionic strengths and pHs. Thermal effects of the difference spectra were cancelled by comparing with the spectra of model compounds such as lysozyme and ribonuclease, and the blue shift of α_s-casein spectra was observed at above 30°C in these all experimental conditions. This shift was considered to mean unfolding of the α_s-casein molecule. The aggregation of α_s-casein was observed above ionic strength of 0.4 by heating. These heat-induced changes were reversible until the aggregation was observed.     

Sex pheromone components of the leafroller moth Pandemis cerasana

Female-tip washings of the leafroller moth, Pandemis cerasana, were found to contain (E)-11-tetradecenyl acetate, (Z)-11-tetradecenyl acetate, (E)-11-tetradecenol, (Z)-11-tetradecenol and tetradecyl acetate, based on chemical analysis and electroantennogram tests. The relative amounts of these compounds in the gland were ca. 64:21:10:3:2 in the order named. Only (E)-11 and (Z)-11-tetradecenyl acetate were required for attraction of males to trap dispensed in the ratio 3:1, respectively.     

A two-component sex attractant for males of the geometrid moth Alsophila quadripunctata

The binary mixture of the hydrocarbons (3Z,6Z,9Z)-3,6,9-nonadecatriene and (6Z,9Z)-6,9-nonadecadiene is a sex attractant for males of Alsophila quadripunctata Esp. (Lepidoptera, Geometridae). Triene/diene ratios from 1:1 to 1:0.01 attracted males in the field, the ratios 1:1 and 1:0.7 were most effective. The triene on its own also attracted males, but in significantly lower numbers than the 1:1 mixture. In a dosage test the highest dose, 1800 g of 1:0.8 [triene:diene] mixture applied to rubber caps was most effective. This is the first report of a geometrid sex attractant with field activity containing more than one component.

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Zusammenfassung Die binäre Mischung der Kohlenwasserstoffe (3Z,6Z,9Z)-3,6,9-Nonadecatrien [ZZZ3,6,9-19:H] und (6Z,9Z)-6,9-Nonadecadien [ZZ6,9-19:H] in den Verhältnissen 1:1 bis 1:0.01 erwies sich im Freiland als Sexuallockstoff für Männchen von Alsophila quadripunctata (Geometridae, Lepidoptera). Dabei besaß die 1:1-Mischung die beste Fängigkeit, das Nonadecatrien allein lockte schlechter, während das C₁₉-Alkadien unwirksam war. Ein Dosiswirksamkeitstest ergab bei Köderbeladungen von 1.8 mg (Trien:Dien + 1:0.8) die besten Fangergebnisse. Im Verlauf der Untersuchungen wurden weitere zwölf geometrische-oder positionsisomere C₁₉-Polyenkohlenwasserstoffe getestet, wobei keine der geprüften Verbindungen Attraktant-oder Inhibitorwirkung zeigte.

     

A Selective Method of Estimation of Bacitracin in the Presence of Other Antibiotics and Growth Factors

The plum cankerworm moth, Cystidia couaggaria couaggaria (Geometridae: Ennominae), is a defoliator of Chinese plum trees (Prunus mume). The pheromone components of the female were analyzed by gas chromatography (GC) with an electro-antennographic (EAG) detector and GC coupled with mass spectrometry. The crude pheromone extract included several EAG-active components, i.e., trienyl, dienyl, and saturated hydrocarbons, with a C₂₁–C₂₅ straight chain. The characteristic mass spectra indicated the unsaturated hydrocarbons to be (3Z,6Z,9Z)-3,6,9-trienes and (6Z,9Z)-6,9-dienes. In the fields, mixtures of the synthetic C₂₁ and C₂₃ trienes in a ratio of 2:3 and 1:4 successfully attracted males of this diurnal species during daytime. While the male antennae responded to the C₂₅ triene and saturated hydrocarbons, their synergistic effects were not observed on the male attraction in the fields. Addition of the C₂₁ diene interestingly inhibited the activity of the triene mixture. Males of Cystidia truncangulata, a sympatric diurnal congener of C. c. couaggaria, showed similar EAG responses to the unsaturated hydrocarbons, but no C. truncangulata males were attracted by the lures tested for C. c. couaggaria males, indicating that the identified hydrocarbons comprised the species-specific pheromone of C. c. couaggaria females.     

Sex pheromone of the butterbur borer, Ostrinia zaguliaevi

The sex pheromone blend of the butterbur borer, Ostrinia zaguliaevi (Lepidoptera: Pyralidae) was analyzed by means of gas chromatography-electroantennographic detection (GC-EAD), GC-mass spectrometry and a series of wind-tunnel bioassays. Four EAD-active compounds were detected in the female sex pheromone gland extract, and these were identified as tetradecyl acetate (14:OAc), (Z)-9-tetradecenyl acetate (Z9-14:OAc), (E)-11-tetradecenyl acetate (E11-14:OAc) and (Z)-11-tetradecenyl acetate (Z11-14:OAc). The average amounts ± s.d. of the four compounds in a single sex pheromone gland were 7.9±3.7 ng, 10.1±3.2 ng, 1.1±0.5 ng and 11.6±5.1 ng, respectively. In a wind-tunnel bioassay, the ternary blend of Z9-, E11- and Z11-14:OAc at a ratio found in the sex pheromone gland (45:5:50) elicited the same behavioral responses from the males as did virgin females and pheromone gland extract. Removal of any single compound from the ternary blend significantly diminished the pheromonal activity, whereas addition of 14:OAc to the ternary blend had no effect on the males' behavioral responses. Therefore, it was concluded that the sex pheromone blend of O. zaguliaevi is composed of Z9-14:OAc, E11-14:OAc and Z11-14:OAc at a ratio of 45:5:50.     

SEX PHEROMONE COMPONENTS OF THE OREIENTAL TOBACCO BUDWORM, HELICOVERPA ASSULTA GUENÉE: IDENTIFICATION AND FIELD TRIALS

Abstract  The sex pheromone gland extracts of the Oriental tobacco budworm, Helicover assulta Guenée, collected from North China in Beijing area, were analyzed by capillary gas chromatograph(GC) and 6 components from the extracts were identified by capillary GC—MS as hexadecanal (16:Ald), (Z)9-hexadecenal (Z9–16:Ald), (Z)-11-hexadecenal (Z11–16:Ald), (Z)-9-hexadecen-1-ol (Z9–16:OH), Z11-hexadecen-1-ol (Z11–16:OH), Z-9-hexadecenylacetate (Z9–16:OAc) at a ratio of 10. 9: 58. 7: 3. 9: 14. 7: 1.1: 10. 7. Field studies indicated that an optimum blend of Z9–16:Ald and Z11–16:Ald was 100: 7. Addition of 16: Ald or Z9–16:Ac to the two aldehyde blend showed no significant effect on attractiveness. However presence of Z9–16:OH in the blends significantly reduced male captures.     

(11Z)-hexadec-11-enal enhances the attractiveness of Diatraea saccharalis main pheromone component in wind tunnel experiments

Abstract:  GC-EAD and GC-MS analysis of pheromone gland extracts of sugarcane borer, Diatraea saccharalis , revealed two antennally active compounds, (9 Z ,11 E )-hexadeca-9,1-dienal and (11 Z )-hexadec-11-enal, in approximately 10 : 1 ratio. Various doses of identified compounds were investigated in wind tunnel experiments individually and in a 10 : 1 ratio. At all tested doses (9 Z ,11 E )-hexadeca-9,1-dienal alone elicited upwind orientation and source location only in a minority of tested males. An admixture of (11 Z )-hex-11-enal enhanced the attractiveness of (9 Z ,11 E )-hexadeca-9,11-dienal significantly. This two-component blend (100 pg) was as attractive as natural pheromone extracted from three female pheromone glands. The data suggest that (11 Z )-hexadec-11-enal is a part of the D. saccharalis sex pheromone.     

Electroantennogram, wind tunnel and field responses of male Mediterranean corn borer, Sesamia nonagrioides, to several blends of its sex pheromone components

The electrophysiological and behavioural responses of male Sesamia nonagrioides Lefèbvre (Lepidoptera: Noctuidae) to several synthetic blends of its sex pheromone components were investigated. Electroantennogram (EAG) recordings showed the greatest response to a 77:8:10:5 blend of the four compounds found in the female glands: (Z)-11-hexadecen-1-yl acetate, (Z)-11-hexadecen-1-ol, (Z)-11-hexadecenal and dodecyl acetate. A logistic equation was fitted to the dose-response data and differences in relative EAG responses were analysed by deriving five parameters of this equation. Flight responses of males were also evaluated in a wind tunnel where the proposed blend (77:8:10:5) also elicited the best response. Field tests confirmed the laboratory findings as the 77:8:10:5 blend captured more S. nonagrioides males than other pheromone mixtures and showed more selectivity for male S. nonagrioides over other noctuid species, Discestra trifolii Rottenberg and Lacanobia oleracea L., but not for Mythimna unipuncta Haworth, a species with a sex pheromone composition almost identical to that of S. nonagrioides.     

Structural basis for pH-dependent alterations of reaction specificity of vertebrate lipoxygenase isoforms

Lipoxygenases have been classified according to their specificity of fatty acid oxygenation and for several plant enzymes pH-dependent alterations in the product patterns have been reported. Assuming that the biological role of mammalian lipoxygenases is based on the formation of specific reaction products, pH-dependent alterations would impact enzymes' functionality. In this study we systematically investigated the pH-dependence of vertebrate lipoxygenases and observed a remarkable stability of the product pattern in the near physiological range for the wild-type enzyme species. Site-directed mutagenesis of selected amino acids and alterations in the substrate concentrations induced a more pronounced pH-dependence of the reaction specificity. For instance, for the V603H mutant of the human 15-lipoxygenase-2 8-lipoxygenation was dominant at acidic pH (65%) whereas 15-H(p)ETE was the major oxygenation product at pH 8. Similarly, the product pattern of the wild-type mouse 8-lipoxygenase was hardly altered in the near physiological pH range but H604F exchange induced strong pH-dependent alterations in the positional specificity. Taken together, our data suggest that the reaction specificities of wild-type vertebrate lipoxygenase isoforms are largely resistant towards pH alterations. However, we found that changes in the assay conditions (low substrate concentration) and introduction/removal of a critical histidine at the active site impact the pH-dependence of reaction specificity for some lipoxygenase isoforms.     

Female sex pheromone of a lichen moth Eilema japonica (Arctiidae, Lithosiinae): Components and control of production

Seven candidates for components of the female sex pheromone of Eilema japonica (Arctiidae, Lithosiinae) were detected in an extract of pheromone glands with a gas chromatograph-electroantennographic detector. The compounds were identified as (Z,Z)-6,9-icosadiene (D20), (Z,Z)-6,9-henicosadiene (D21), (Z,Z,Z)-3,6,9-henicosatriene (T21), (Z,Z)-6,9-docosadiene (D22), (Z,Z,Z)-3,6,9-docosatriene (T22), (Z,Z)-6,9-tricosadiene (D23), and (Z,Z,Z)-3,6,9-tricosatriene (T23). Assays using synthetic lures in a wind tunnel showed that D21 (proportion, 0.39), T21 (0.08), D22 (0.27), and T22 (0.26) are important for evoking full behavioral responses from the males. Titers of the pheromone components did not show clear temporal fluctuations. Moreover, decapitation of the female moth had no effect on the titers of pheromone components in the pheromone gland, suggesting that cephalic endocrine factors such as pheromone biosynthesis activating neuropeptide (PBAN) are not involved in the control of pheromone biosynthesis in this species.     

Role of Arg403 for thermostability and catalytic activity of rabbit 12/15-lipoxygenase

12/15-Lipoxygenases (12/15-LOX) have been implicated in inflammatory and hyperproliferative diseases but the numerous aspects of structural biology of these enzymes are far from clear. Early mutagenesis data and structural modeling of enzyme–substrate complexes suggested that Arg403, which is localized at the entrance of the putative substrate binding pocket, might interact with the fatty acid carboxylic group. On the other hand, side-chain of Arg403 is a part of an ionic network with the residues of α2-helix, which undergoes pronounced conformation changes upon inhibitor binding. To explore the role of Arg403 for catalysis in more detail we exchanged positively charged Arg403 to neutral Leu and quantified structural and functional consequences of the alteration at the site of mutation using fluorometric techniques. We found that a loss of electrostatic interaction between Arg403 and negatively charged amino acid residues of α2-helix has only minor impact on protein folding, but partially destabilized the tertiary structure of the enzyme. We hypothesize that interaction of Arg403 with the substrate's carboxylate might be involved in a complex mechanism triggering conformational changes of the α2-helix, which are required for formation of the catalytically competent dimer r12/15-LOX complex at pre-catalytic stages.