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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Photosynthesis research 38 (1993), S. 441-448 
    ISSN: 1573-5079
    Keywords: hydroxylamine ; hypochlorite ; manganese ; oxygen evolution ; Photosystem II ; S-states
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Treatment of Photosystem II (PS II) with low concentrations of hydroxylamine is known to cause a two-flash delay in the O2-evolution pattern, and in the formation of the S2-state multiline EPR signal, due to the two-electron reduction of the S1-state by hydroxylamine to form the S-1-state. Past work has shown that these delays are not reversed by washing out the hydroxylamine nor by adding DCBQ or ferricyanide to oxidize the residual hydroxylamine, but are reversed by illumination with two saturating flashes followed by a 30-min dark incubation. We have examined the effects of treatments aimed at restoring the normal flash-induced O2-evolution pattern and S2-state multiline EPR signal after treatment of PS II with 40 μM hydroxylamine. In agreement with past work, we find that the two-flash delay in O2 evolution is not reversed when the hydroxylamine is removed by three cycles of centrifugation and resuspension in hydroxylamine-free buffer nor by adding ferricyanide or DCBQ to oxidize the unreacted hydroxylamine. However, the normal flash-induced O2-evolution pattern is restored by illumination with two saturating flashes followed by a 30-min dark incubation (after the sample was first treated with 40 μM hydroxylamine and the unreacted hydroxylamine was removed); illumination with one saturating flash followed by a 30-min dark incubation is only partially effective. These results show that ferricyanide and DCBQ are not effective at oxidizing the S-1-state to the S1-state. In contrast, adding hypochlorite (OCl-) after treatment with hydroxylamine restored the normal flash-induced O2-evolution pattern and also restored the formation of the S2-state multiline EPR signal by illumination at 200 K. We conclude that hypochlorite is capable of oxidizing the S-1-state to the S1-state. This is the first example of a chemical treatment that advances the delayed flash-induced O2 evolution pattern.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biospectroscopy 2 (1996), S. 167-171 
    ISSN: 1075-4261
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Physics
    Notes: Fluorescence measurements of photosynthetic organisms and isolated proteins at ambient and low temperature have played an important role in understanding their function. When comparing fluorescence measurements at cryogenic temperatures, the differences in the scattering properties of frozen samples make it difficult to compare the fluorescence intensity of these samples. An internal emission standard can be used to scale the fluorescence intensity, compensating for these differences. We report the synthesis, purification and characterization of a luminescent terbium chelate complex for use as an internal emission standard for the study of photosystem II fluorescence at cryogenic temperatures. The ligand consists of a diethylenetriaminepentaacetic acid derivative where pyrimidine rings sensitize the terbium luminescence, overcoming the inherently low absorption of terbium. The chelated lanthanide remains in solution in the aqueous phase and does not interfere with photosystem II function. By scaling to the terbium emission, the fluorescence intensity of different samples can be readily compared. This chelate complex could also be used as an internal emission standard for studies of other proteins. © 1996 John Wiley & Sons, Inc.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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