title: Concentration-dependent oligomerization of cross-linked complexes between ferredoxin and ferredoxin-NADP+ reductase.
abstract: Ferredoxin-NADP(+) reductase (FNR) forms a 1:1 complex with ferredoxin (Fd), and catalyzes the electron transfer between Fd and NADP(+). In our previous study, we prepared a series of site-specifically cross-linked complexes of Fd and FNR, which showed diverse electron transfer properties. Here, we show that X-ray crystal structures of the two different Fd-FNR cross-linked complexes form oligomers by swapping Fd and FNR moieties across the molecules; one complex is a dimer from, and the other is a successive multimeric form. In order to verify whether these oligomeric structures are formed only in crystal, we investigated the possibility of the oligomerization of these complexes in solution. The mean values of the particle size of these cross-linked complexes were shown to increase with the rise of protein concentration at sub-milimolar order, whereas the size of dissociable wild-type Fd:FNR complex was unchanged as analyzed by dynamic light scattering measurement. The oligomerization products were detected by SDS-PAGE after chemical cross-linking of these complexes at the sub-milimolar concentrations. The extent and concentration-dependent profile of the oligomerizaion were differentiated between the two cross-linked complexes. These results show that these Fd-FNR cross-linked complexes exhibit concentration-dependent oligomerization, possibly through swapping of Fd and FNR moieties also in solution. These findings lead to the possibility that some native multi-domain proteins may present similar phenomenon in vivo.
authors.affiliation: Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan. a-yoko@protein.osaka-u.ac.jp
authors.forename: Yoko
authors.initials: Y
authors.lastname: Kimata-Ariga
authors.forename: Hisako
authors.initials: H
authors.lastname: Kubota-Kawai
authors.forename: Young-Ho
authors.initials: YH
authors.lastname: Lee
authors.forename: Norifumi
authors.initials: N
authors.lastname: Muraki
authors.forename: Takahisa
authors.initials: T
authors.lastname: Ikegami
authors.forename: Genji
authors.initials: G
authors.lastname: Kurisu
authors.forename: Toshiharu
authors.initials: T
authors.lastname: Hase
mesh_terms: D003432:Cross-Linking Reagents / Q000737:chemistry; D018360:Crystallography, X-Ray; D004579:Electron Transport; D004591:Electrophoresis, Polyacrylamide Gel; D005287:Ferredoxin-NADP Reductase / Q000737:chemistry* / Q000378:metabolism; D005288:Ferredoxins / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D046912:Multiprotein Complexes / Q000737:chemistry* / Q000378:metabolism; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary
chemical_list: D003432:Cross-Linking Reagents; D005288:Ferredoxins; D046912:Multiprotein Complexes; D005287:Ferredoxin-NADP Reductase