Structure of the cytosolic part of the subunit b-dimer of Escherichia coli F0F1-ATP synthase.
The structure of the external stalk and its function in the catalytic mechanism of the F(0)F(1)-ATP synthase remains one of the important questions in bioenergetics. The external stalk has been proposed to be either a rigid stator that binds F(1) or an elastic structural element that transmits energy from the small rotational steps of subunits c to the F(1) sector during catalysis. We employed proteomics, sequence-based structure prediction, molecular modeling, and electron spin resonance spectroscopy using site-directed spin labeling to understand the structure and interfacial packing of the Escherichia coli b-subunit homodimer external stalk. Comparisons of bacterial, cyanobacterial, and plant b-subunits demonstrated little sequence similarity. Supersecondary structure predictions, however, show that all compared b-sequences have extensive heptad repeats, suggesting that the proteins all are capable of packing as left-handed coiled-coils. Molecular modeling subsequently indicated that b(2) from the E. coli ATP synthase could pack into stable left-handed coiled-coils. Thirty-eight substitutions to cysteine in soluble b-constructs allowed the introduction of spin labels and the determination of intersubunit distances by ESR. These distances correlated well with molecular modeling results and strongly suggest that the E. coli subunit b-dimer can stably exist as a left-handed coiled-coil.
Hornung
Tassilo
T
Department of Biological Sciences, Southern Methodist University, Dallas, Texas, USA.
Volkov
Oleg A
OA
Zaida
Tarek M A
TM
Delannoy
Sabine
S
Wise
John G
JG
Vogel
Pia D
PD
D003198:Computer Simulation; D003600:Cytosol / Q000737:chemistry*; D019281:Dimerization; D004926:Escherichia coli / Q000201:enzymology*; D007536:Isomerism; D008956:Models, Chemical; D008958:Models, Molecular; D011487:Protein Conformation; D021122:Protein Subunits; D006180:Proton-Translocating ATPases / Q000737:chemistry* / Q000648:ultrastructure*
D021122:Protein Subunits; D006180:Proton-Translocating ATPases