FAD assembly and thylakoid membrane binding of ferredoxin:NADP+ oxidoreductase in chloroplasts.
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.
Onda
Yayoi
Y
Division of Enzymology, Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan. enzyme@protein.osaka-u.ac.jp
Hase
Toshiharu
T
D002736:Chloroplasts / Q000378:metabolism*; D005287:Ferredoxin-NADP Reductase / Q000096:biosynthesis* / Q000378:metabolism*; D005288:Ferredoxins / Q000378:metabolism; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism*; D008565:Membrane Proteins / Q000096:biosynthesis / Q000378:metabolism; D018515:Plant Leaves / Q000166:cytology; D010940:Plant Proteins / Q000096:biosynthesis / Q000378:metabolism; D018724:Spinacia oleracea / Q000166:cytology; D020524:Thylakoids / Q000378:metabolism*; D003313:Zea mays / Q000737:chemistry
D005288:Ferredoxins; D008565:Membrane Proteins; D010940:Plant Proteins; D005182:Flavin-Adenine Dinucleotide; D005287:Ferredoxin-NADP Reductase