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Spectral characterization in the uv region of the precursor of photosynthetically evolved oxygen in isolated trypsinized chloroplasts

, : Spectral characterization in the uv region of the precursor of photosynthetically evolved oxygen in isolated trypsinized chloroplasts. Biochimica et Biophysica Acta 722(1): 1-11

The reaction mechanism of the water-spliting enzyme system Y was analyzed by measurements of UV absorption changes induced by a flash train in dark-adapted spinach chloroplasts. To eliminate processes which give rise to interfering absorption changes with a binary oscillation, mildly trypsizined chloroplasts were applied which are disconnected from the secondary plastoquinone acceptor B, but are able to evolve O2, provided that K3Fe(CN)6 is used as electron acceptor. The extent of the relaxation kinetics of the 320 nm absorption change accomplished within 5 ms oscillates with a periodicity of 4 in phase with the O2 yield as a function of the number of the flash in the train. The difference spectra of the difference between the initial amplitudes and those remaining after a 4.5 ms dark period, referred to as .**GRAPHIC**. exhibit pronounced bands peaking around 320 nm (positive) and 270 nm (negative). The extent oscillates in the flash train. It is at a maximum for the absorption change differences induced by the 3rd flash, .**GRAPHIC**. Taking into account the fact that the 1st flash of the train does not yield O2, whereas the O2 production is at a maximum due to the 3rd flash, the difference of .**GRAPHIC**. as a function of wavelength is interpreted as the difference spectrum of the precursor of molecular O2 in system Y. The spectrum exhibits a band peaking around 320 nm. The precursor is referred to as Y-320. A molecular model proposed for the mechanism of photosynthetic water oxidation is discussed.

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