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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 40 (1999), S. 997-1008 
    ISSN: 1573-5028
    Keywords: glutathione transferase ; heterodimer ; recombinant enzyme ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two cDNAs encoding novel type III maize (Zea mays) GST subunits, ZmGST VI and ZmGST VII, have been cloned in addition to the previously described ZmGST V. Together with the type I GSTs ZmGST I and ZmGST III, these subunits were expressed in Escherichia coli, both individually and in tandem combinations using a customised pET vector. The GST dimers formed were then characterised. When type I GSTs were co-expressed only the respective homodimers were formed rather than the ZmGST I-III heterodimer. The failure to form this heterodimer, together with the negligible herbicide-detoxifying activity associated with recombinant ZmGST III-III, suggests that the identity of herbicide-detoxifying isoenzymes described in maize as being composed of ZmGST III subunits requires re-evaluation. In contrast, co-expression of the type III GSTs ZmGST V and ZmGST VI resulted in the formation of ZmGST V-V, ZmGST VI-VI and ZmGST V-VI dimers in the ratio 1:1:2 as predicted for random subunit association. ZmGST V-VI had kinetic characteristics intermediate between those of the two homodimers, indicating that the subunits were catalytically independent of one another. Co-expression of ZmGST V and ZmGST VII resulted in the formation of ZmGST V-VII and this isoenzyme was subsequently identified in maize plants. Attempts to dimerise type I GST subunits with type III GST subunits proved unsuccessful. These results demonstrate the utility of co-expressing recombinant GSTs to explore the potential of subunit-subunit associations and to help unravel the complexity of homodimeric and heterodimeric GSTs in plants.
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  • 2
    ISSN: 1573-5028
    Keywords: Zea mays ; glutathione transferase ; herbicide ; herbicide safener
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The glutathione transferases (GSTs) from maize (Zea mays L.) with activities toward the chloroacetanilide herbicide metolachlor and the diphenyl ether herbicide fluorodifen were fractionated into two pools based on binding to affinity columns. Pool 1 GSTs were retained on Orange A agarose and were identified as isoenzymes Zea mays (Zm) GST I-I, Zm GST I-II and Zm GST I-III, which have been described previously. Pool 2 GSTs selectively bound to S-hexyl-glutathione-Sepharose and were distinct from the pool 1 GSTs, being composed of a homodimer of 28.5 kDa subunits, termed Zm GST V-V, and a heterodimer of the 28.5 kDa polypeptide and a 27.5 kDa subunit, termed Zm GST V-VI. Using an antibody raised to Zm GST V-VI, a cDNA expression library was screened and a Zm GST V clone identified showing sequence similarity to the type-III auxin-inducible GSTs previously identified in tobacco and other dicotyledenous species. Recombinant Zm GST V-V showed high GST activity towards the diphenyl ether herbicide fluorodifen, detoxified toxic alkenal derivatives and reduced organic hydroperoxides. Antibodies raised to Zm GST I-II and Zm GST V-VI were used to monitor the expression of GST subunits in maize seedlings. Over a 24 h period the Zm GST I subunit was unresponsive to chemical treatment, while expression of Zm GST II was enhanced by auxins, herbicides, the herbicide safener dichlormid and glutathione. The Zm GST V subunit was more selective in its induction, only accumulating significantly in response to dichlormid treatment. During development Zm GST I and Zm GST V were expressed more in roots than in shoots, with Zm GST II expression limited to the roots.
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  • 3
    ISSN: 1040-7685
    Keywords: Phase behavior ; supercritical fluid chromatography ; supercritical fluid extraction ; modifier ; propylene carbonate ; critical parameters ; Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The phase behavior (P-T-x) of propylene carbonate was studied using a variable volume view cell for the regions applicable to supercritical fluid chromatography (SFC) and extraction (SFE). Type 5 phase behavior was observed. Phase separation was shown to significantly alter retention and selectivity in the analysis of coal extracts by capillary SFC. Propylene carbonate was found to have limited use as a modifier in capillary SFC. With 5.1 mol% propylene carbonate at 60°C, a single phase is maintained only at pressures above 250 atm.Twenty-two methods used to estimate critical pressure (Pc), twenty-seven methods used to estimate critical temperature (Tc), and thirty methods used to estimate critical volume (Vc) were evaluated for various modifiers employed in supercritical fluid chromatography (SFC). None of the methods could predict Pc, Tc, or Vc to within a 5% relative error for all of the modifiers tested. However, several of the methods were successful in predicting Pc, Tc, or Vc to within 5% for select chemical classes of compounds.Four methods used to estimate the vapor-liquid critical loci of binary mixtures not only failed to accurately predict vapor-liquid separation, but failed to predict the type of phase separation occurring. None of the above methods could reliably estimate the critrical parameters of propylene carbonate or the critical parameters of the mixture.
    Additional Material: 5 Ill.
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  • 4
    Electronic Resource
    Electronic Resource
    Chichester : Wiley-Blackwell
    Biological Mass Spectrometry 20 (1985), S. 115-121 
    ISSN: 0030-493X
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The positive ion-molecule reactions of OCS have been investigated in an ion cyclotron resonance spectrometer. A variety of reactions in OCS/hydrocarbon mixtures have been investigated for various C1—C4 hydrocarbons - alkanes, alkenes and alkynes. The formation of organosulfur ions is found in reactions in OCS/hydrocarbon (Cn) mixtures with n 〈4. Formation of organosulfur ions is observed from hydrocarbon ions reacting with OCS and [OCS]+· and S+· reacting with the hydrocarbons. The proton affinity of OCS has been determined to be 688.7±8 kJ mol-1 while that of CS2 is measured to be 712.1±8 kJ mol-1. Comparison with the proton affinity of CO2 shows that the proton affinity increases as sulfur is substituted for oxygen.
    Additional Material: 7 Tab.
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