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  • 1
    ISSN: 1520-4995
    Source: ACS Legacy Archives
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-5079
    Keywords: chlorophyll a fluorescence ; oxygen evolving PS II particles ; pheophytin ; photoinhibition ; photosystem II ; primary electron acceptor QA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Oxygen evolving photosystem II particles were exposed to 100 and 250 W m−2 white light at 20°C under aerobic, anaerobic and strongly reducing (presence of dithionite) conditions. Three types of photoinactivation processes with different kinetics could be distinguished: (1) The fast process which occurs under strongly reducing (t 1/2≅1–3 min) and anaerobic conditions (t 1/2≅4–12 min). (2) The slow process (t 1/2≅15–40 min) and (3) the very slow process (t 1/2〉100 min), both of which occur under all three sets of conditions. The fast process results in a parallel decline of variable fluorescence (F v) and of Hill reaction rate, accompanied by an antiparallel increase of constant fluorescence (F o). We assume that trapping of QA in a negatively charged stable state, (QA −)stab, is responsible for the effects observed. The slow process is characterized by a decline of maximal fluorescence (F m). In presence of oxygen this decline is due to the well known disappearance of F v which proceeds in parallel with the inhibition of the Hill reaction; F o remains essentially constant. Under anaerobic and reducing conditions the decline of F m represents the disappearance of the increment in F o generated by the fast process. We assume that the slow process consists in neutralization of the negative charge in the domain of QA in a manner that renders QA non-functional. The charge separation in the RC is still possible, but energy of excitation becomes thermally dissipated. The very slow photoinactivation process is linked to loss of charge separation ability of the PS II RC and will be analyzed in a forthcoming paper.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-5079
    Keywords: Photosystem II ; reaction centers ; Synechococcus ; IMAC ; Cu2+ loaded Sepharose ; QA binding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Oxygen-evolving PS II particles from the thermophilic cyanobacterium Synechococcus elongatus are partially purified by centrifugation on a sucrose gradient and are bound to a Chelating Sepharose column loaded with Cu2+ ions. Bound particles are then transformed into PS II RC complexes by two washing steps. First, washing with a phosphate buffer (pH=6.5) containing 0.02% of SB 12 removes the rest of phycobilins and leaves pure PS II core particles on the column. Second, washing with a phosphate buffer (pH=6.2) containing 0.2 M LiClO4 and 0.05% of DM removes CP 47 and CP 43 and leaves bare PS II RC complexes on the column. These are then eluted with a phosphate buffer containing 1% of dodecylmaltoside (DM). The molar ratio of pigments in the eluate changes with the progress of elution but around the middle of the elution period a nearly stable ratio is maintained of Chl a: Pheo a: Car: Cyt b 559 equal to 2.9: 1: 0.9: 0.8. In these fractions the photochemical separation of charges could be demonstrated by accumulation of reduced pheophytin (ΔA of 430–440 nm) and by the flash induced formation of P680+ (ΔA at 820 nm). The relatively slow relaxation kinetics of the latter signal (t1/2 ≈ 1 ms) may suggest that in a substantial fraction of the RCs QA remains bound to the complex.
    Type of Medium: Electronic Resource
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