engleski

Importance of ferredoxin:NADP+ oxidoreductase in photosynthetic alternative electron transfer pathways

Ferredoxin:NADP+ oxidoreductase (FNR) catalyses reversible reaction of electron transfer from ferredoxin to NADP+ in photosynthesis. FNR is present as a soluble protein in stroma, but also as dynamically bound to the protein complexes with the thylakoid rhodanase-like protein (TROL) and the translocon on the inner envelope chloroplast membrane 62 (Tic62). It is envisaged that ferredoxin efficiently passes electrons to alternative pathways when FNR is not sequestered to the TROL. We are using the common C3 model plant organism Arabidopsis thaliana L. Col-0 that has two leaf FNR isoforms and Zea mays, a C4 plant that contains three isoforms. We have chosen maize FNR1 because it has been shown that ZmFNR1 strongly binds to the TROL. FLAG and HA tags were PCR-added to the 3´-end of the ZmFNR1 gene to enable subsequent isolation and analyses of protein complexes via tandem affinity purification. By using Gateway cloning technology, we have constructed Arabidopsis transgenic plants containing ZmFNR1 bearing C-terminal FLAG and HA tags. By crossing those plants with the Arabidopsis plants that accumulate TROL with amino acid or domain changes we have created specific experimental platforms which will be used to explore TROL structures that are involved in the dynamic FNR binding under different light conditions.

photosynthesis ; FNR ; TROL ; Arabidopsis thaliana ; protein complexes

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