X-ray scattering studies of Methylophilus methylotrophus (sp. W3A1) electron-transferring flavoprotein. Evidence for multiple conformational states and an induced fit mechanism for assembly with trimethylamine dehydrogenase.
Small angle x-ray solution scattering has been used to generate a low resolution, model-independent molecular envelope structure for electron-transferring flavoprotein (ETF) from Methylophilus methylotrophus (sp. W(3)A(1)). Analysis of both the oxidized and 1-electron-reduced (anionic flavin semiquinone) forms of the protein revealed that the solution structures of the protein are similar in both oxidation states. Comparison of the molecular envelope of ETF from the x-ray scattering data with previously determined structural models of the protein suggests that ETF samples a range of conformations in solution. These conformations correspond to a rotation of domain II with respect to domains I and III about two flexible "hinge" sequences that are unique to M. methylotrophus ETF. The x-ray scattering data are consistent with previous models concerning the interaction of M. methylotrophus ETF with its physiological redox partner, trimethylamine dehydrogenase. Our data reveal that an "induced fit" mechanism accounts for the assembly of the trimethylamine dehydrogenase-ETF electron transfer complex, consistent with spectroscopic and modeling studies of the assembly process.
Jones
M
M
Council for the Central Laboratory of the Research Councils Daresbury Laboratory, Warrington, Cheshire WA4 4AD, United Kingdom.
Basran
J
J
Sutcliffe
M J
MJ
Günter Grossmann
J
J
Scrutton
N S
NS
D003001:Cloning, Molecular; D004579:Electron Transport; D044927:Electron-Transferring Flavoproteins; D004926:Escherichia coli; D005420:Flavoproteins / Q000737:chemistry* / Q000378:metabolism; D020630:Methylophilus methylotrophus / Q000378:metabolism*; D008958:Models, Molecular; D010089:Oxidoreductases, N-Demethylating / Q000378:metabolism*; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012542:Scattering, Radiation; D013053:Spectrophotometry; D014965:X-Rays
D044927:Electron-Transferring Flavoproteins; D005420:Flavoproteins; D011994:Recombinant Proteins; D010089:Oxidoreductases, N-Demethylating; C023186:trimethylamine dehydrogenase