Electron transfer of site-specifically cross-linked complexes between ferredoxin and ferredoxin-NADP(+) reductase.
Ferredoxin (Fd) and Fd-NADP(+) reductase (FNR) are redox partners responsible for the conversion between NADP(+) and NADPH in the plastids of photosynthetic organisms. Introduction of specific disulfide bonds between Fd and FNR by engineering cysteines into the two proteins resulted in 13 different Fd-FNR cross-linked complexes displaying a broad range of activity to catalyze the NADPH-dependent cytochrome c reduction. This variability in activity was thought to be mainly due to different levels of intramolecular electron transfer activity between the FNR and Fd domains. Stopped-flow analysis revealed such differences in the rate of electron transfer from the FNR to Fd domains in some of the cross-linked complexes. A group of the cross-linked complexes with high cytochrome c reduction activity comparable to dissociable wild-type Fd/FNR was shown to assume a similar Fd-FNR interaction mode as in the native Fd:FNR complex by analyses of NMR chemical shift perturbation and absorption spectroscopy. However, the intermolecular electron transfer of these cross-linked complexes with two Fd-binding proteins, nitrite reductase and photosystem I, was largely inhibited, most probably due to steric hindrance by the FNR moiety linked near the redox center of the Fd domain. In contrast, another group of the cross-linked complexes with low cytochrome c reduction activity tends to mediate higher intermolecular electron transfer activity. Therefore, reciprocal relationship of intramolecular and intermolecular electron transfer abilities was conferred by the linkage of Fd and FNR, which may explain the physiological significance of the separate forms of Fd and FNR in chloroplasts.
Laboratory of Regulation of Biological Reactions, Institute for Protein Research,Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan. a-yoko@protein.osaka-u.ac.jp
Yoko
Y
Kimata-Ariga
Yukiko
Y
Sakakibara
Takahisa
T
Ikegami
Toshiharu
T
Hase
D002384:Catalysis; D004579:Electron Transport; D014450:Electron Transport Complex III / Q000737:chemistry / Q000378:metabolism; D005287:Ferredoxin-NADP Reductase / Q000737:chemistry* / Q000378:metabolism; D005288:Ferredoxins / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular
D005288:Ferredoxins; D005287:Ferredoxin-NADP Reductase; D014450:Electron Transport Complex III