title: Ferredoxin:NADP+ oxidoreductase is a subunit of the chloroplast cytochrome b6f complex.
abstract: Purified detergent-soluble cytochrome b6f complex from chloroplast thylakoid membranes (spinach) and cyanobacteria (Mastigocladus laminosus) was highly active, transferring 300-350 electrons per cyt f/s. Visible absorbance spectra showed a red shift of the cytochrome f alpha-band and the Qy chlorophyll a band in the cyanobacterial complex and an absorbance band in the flavin 450-480-nm region of the chloroplast complex. An additional high molecular weight (M(r) approximately 35,000) polypeptide in the chloroplast complex was seen in SDS-polyacrylamide gel electrophoresis at a stoichiometry of approximately 0.9 (cytochrome f)(-1). The extra polypeptide did not stain for heme and was much more accessible to protease than cytochrome f. Electrospray ionization mass spectrometry of CNBr fragments of the 35-kDa polypeptide was diagnostic for ferredoxin:NADP+ oxidoreductase (FNR), as were antibody reactivity to FNR and diaphorase activity. The absence of FNR in the cyanobacterial complex did not impair decyl-plastoquinol-ferricyanide activity. The activity of the FNR in the chloroplast b6f complex was also shown by NADPH reduction, in the presence of added ferredoxin, of 0.8 heme equivalents of the cytochrome b6 subunit. It was inferred that the b6f complex with bound FNR, one equivalent per monomer, provides the membrane protein connection to the main electron transfer chain for ferredoxin-dependent cyclic electron transport.
authors.affiliation: Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907-1392, USA.
authors.forename: H
authors.initials: H
authors.lastname: Zhang
authors.forename: J P
authors.initials: JP
authors.lastname: Whitelegge
authors.forename: W A
authors.initials: WA
authors.lastname: Cramer
mesh_terms: D000595:Amino Acid Sequence; D015153:Blotting, Western; D002736:Chloroplasts / Q000201:enzymology*; D003001:Cloning, Molecular; D003573:Cytochrome b Group / Q000737:chemistry / Q000378:metabolism*; D045346:Cytochrome b6f Complex; D004579:Electron Transport; D004591:Electrophoresis, Polyacrylamide Gel; D005287:Ferredoxin-NADP Reductase / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D017386:Sequence Homology, Amino Acid; D021241:Spectrometry, Mass, Electrospray Ionization; D018724:Spinacia oleracea
chemical_list: D003573:Cytochrome b Group; D045346:Cytochrome b6f Complex; D005287:Ferredoxin-NADP Reductase