Scanning probe microscopic studies of the oriented attachment and membrane reconstitution of cytochrome C oxidase to a gold electrode.
Scanning probe microscopy was used to monitor the resulting surface of the oriented incorporation of cytochrome c oxidase into electrode supported lipid bilayer at four crucial stages with molecular resolution. We were able to reveal the formation of a densely packed monolayer of the active ester dithio(succiniimidylepropionate) (DTSP) and the covalent linkage of the nitrilotriacetic acid (NTA) to the thiol anchored DTSP by scanning tunneling microscopy. Atomic force microscopy investigations showed that the detergent solubilized oxidase is immobilized as monomers and small aggregates via histidine residues. Finally, the reconstitution of the proteins within the supported membrane was verified. The amount of oxidase immobilized within the solid supported membrane was estimated.
Mayer
Dirk
D
Research Center Juelich, ISG-2: Institute for Bio and Chemosensors, IBI-2: Structural Biology, D-52425 Juelich, Germany.
Ataka
Kenichi
K
Heberle
Joachim
J
Offenhäusser
Andreas
A
D004566:Electrodes; D003576:Electron Transport Complex IV / Q000737:chemistry*; D006046:Gold / Q000737:chemistry*; D008567:Membranes, Artificial; D020527:Microscopy, Scanning Probe / Q000379:methods*; D010316:Particle Size; D012680:Sensitivity and Specificity; D013499:Surface Properties
D008567:Membranes, Artificial; D006046:Gold; D003576:Electron Transport Complex IV