FAD assembly and thylakoid membrane binding of ferredoxin:NADP+ oxidoreductase in chloroplasts |
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Authors: | Onda Yayoi Hase Toshiharu |
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Institution: | Division of Enzymology, Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan. enzyme@protein.osaka-u.ac.jp |
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Abstract: | We investigated the process of flavin adenine dinucleotide (FAD) incorporation into the ferredoxin (Fd):NADP(+) oxidoreductase (FNR) polypeptide during FNR biosynthesis, using pull-down assay with resin-immobilized Fd which bound strongly to FAD-assembled holo-FNR, but hardly to FAD-deficient apo-FNR. After FNR precursor was imported into isolated chloroplasts and processed to the mature size, the molecular form pulled down by Fd-resin increasingly appeared. The mature-sized FNR (mFNR) accumulated transiently in the stroma as the apo-form, and subsequently bound on the thylakoid membranes as the holo-form. Thus, FAD is incorporated into the mFNR inside chloroplasts, and this assembly process is followed by the thylakoid membrane localization of FNR. |
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