Laser flash absorption spectroscopy study of flavodoxin reduction by photosystem I in Synechococcus sp. PCC 7002.
The photoreduction of flavodoxin by trimeric photosystem I, both from the cyanobacterium Synechococcus sp. PCC 7002, was investigated by flash absorption spectroscopy. After addition of flavodoxin in darkness, single flash experiments show that the transient signals change between individual flashes. This behavior is assigned to a progressive accumulation of flavodoxin semiquinone, which is relatively stable under most experimental conditions. Different conditions were devised in order to study the reduction of the oxidized and semiquinone forms of flavodoxin separately. Both processes were identified by their differential spectra measured between 460 and 630 nm. Detailed kinetic characteristics of flavodoxin reduction were obtained at pH 8.0 in the presence of salts. The kinetics of reduction of oxidized flavodoxin displays a single-exponential component. The rate of this component increases with the flavodoxin concentration up to an asymptotic value of about 600 s-1. The semiquinone form of flavodoxin being protonated, this rate corresponds to a rate-limiting reaction which could be either an electron transfer reaction or a protonation reaction. In contrast, the reduction of flavodoxin semiquinone is biphasic. A fast first-order phase with t 1/2 approximately 10 microseconds is interpreted as an electron transfer process within a preformed complex. A dissociation constant of 2.64 microM is calculated for this complex by assuming a simple binding equilibrium between photosystem I and flavodoxin semiquinone. The slower phase observed for semiquinone reduction is concentration dependent, and a second-order rate constant of 1.7 x 10(8) M-1 s-1 is calculated. For both one-electron reduction steps, different optimal salt concentrations are observed indicating slightly different interactions between photosystem I and flavodoxin in its oxidized and semiquinone states.
Mühlenhoff
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Institut für Biologie II, Universität Freiburg, FRG.
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D000458:Cyanobacteria / Q000378:metabolism*; D005418:Flavodoxin / Q000378:metabolism*; D007700:Kinetics; D007834:Lasers; D015636:Magnesium Chloride / Q000494:pharmacology; D008956:Models, Chemical; D010084:Oxidation-Reduction / Q000187:drug effects; D045322:Photosynthetic Reaction Center Complex Proteins / Q000187:drug effects / Q000378:metabolism*; D045331:Photosystem I Protein Complex; D012965:Sodium Chloride / Q000494:pharmacology; D013053:Spectrophotometry / Q000379:methods
D005418:Flavodoxin; D045322:Photosynthetic Reaction Center Complex Proteins; D045331:Photosystem I Protein Complex; D015636:Magnesium Chloride; D012965:Sodium Chloride