Structure of functional Staphylococcus aureus alpha-hemolysin channels in tethered bilayer lipid membranes.
We demonstrate a method for simultaneous structure and function determination of integral membrane proteins. Electrical impedance spectroscopy shows that Staphylococcus aureus alpha-hemolysin channels in membranes tethered to gold have the same properties as those formed in free-standing bilayer lipid membranes. Neutron reflectometry provides high-resolution structural information on the interaction between the channel and the disordered membrane, validating predictions based on the channel's x-ray crystal structure. The robust nature of the membrane enabled the precise localization of the protein within 1.1 A. The channel's extramembranous cap domain affects the lipid headgroup region and the alkyl chains in the outer membrane leaflet and significantly dehydrates the headgroups. The results suggest that this technique could be used to elucidate molecular details of the association of other proteins with membranes and may provide structural information on domain organization and stimuli-responsive reorganization for transmembrane proteins in membrane mimics.
McGillivray
Duncan J
DJ
National Institute of Standards and Technology (NIST) Center for Neutron Research, Gaithersburg, Maryland, USA.
Valincius
Gintaras
G
Heinrich
Frank
F
Robertson
Joseph W F
JW
Vanderah
David J
DJ
Febo-Ayala
Wilma
W
Ignatjev
Ilja
I
Lösche
Mathias
M
Kasianowicz
John J
JJ
D001427:Bacterial Toxins / Q000737:chemistry*; D006046:Gold / Q000737:chemistry; D006460:Hemolysin Proteins / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry*; D008958:Models, Molecular; D018408:Patch-Clamp Techniques; D013057:Spectrum Analysis; D013211:Staphylococcus aureus
D001427:Bacterial Toxins; D006460:Hemolysin Proteins; D008051:Lipid Bilayers; C021091:staphylococcal alpha-toxin; D006046:Gold