Photo-induced electron transfer from photosystem I to NADP(+): characterization and tentative simulation of the in vivo environment.
The photoproduction of NADPH in photosynthetic organisms requires the successive or concomitant interaction of at least three proteins: photosystem I (PSI), ferredoxin (Fd) and ferredoxin:NADP(+) oxidoreductase (FNR). These proteins and their surrounding medium have been carefully analysed in the cyanobacterium Synechocystis sp. PCC 6803. A high value of 550mg/ml was determined for the overall solute content of the cell soluble compartment. PSI and Fd are present at similar concentrations, around 500μM, whereas the FNR associated to phycobilisome is about 4 fold less concentrated. Membrane densities of FNR and trimeric PSI have been estimated to 2000 and 2550 per μm(2), respectively. An artificial confinement of Fd to PSI was designed using fused constructs between Fd and PsaE, a peripheral and stroma located PSI subunit. The best covalent system in terms of photocatalysed NADPH synthesis can be equivalent to the free system in a dilute medium. In a macrosolute crowded medium (375mg/ml), this optimized PSI/Fd covalent complex exhibited a huge superiority compared to the free system. This is a likely consequence of restrained diffusion constraints due to the vicinity of two out of the three protein partners. In vivo, Fd is the free partner, but the constant proximity between PSI and the phycobilisome associated FNR creates a similar situation, with two closely associated partners. This organization seems well adapted for an efficient in vivo production of the stable and fast diffusing NADPH.
Service de Bioenergetique, Biologie Structurale et Mecanismes, Gif sur Yvette, France.
Gwenaëlle
G
Moal
Bernard
B
Lagoutte
D004058:Diffusion; D004579:Electron Transport; D005288:Ferredoxins / Q000378:metabolism; D009249:NADP / Q000096:biosynthesis*; D045331:Photosystem I Protein Complex / Q000378:metabolism*; D046939:Synechocystis / Q000378:metabolism*
D005288:Ferredoxins; D045331:Photosystem I Protein Complex; D009249:NADP