Streptavidin binding observed with an atomic force microscope.
An atomic force microscope (AFM) was used to investigate a specific recognition reaction: the binding of streptavidin to a biotinylated lipid bilayer. Prior to the recognition reaction, the phase coexistence of the lipid bilayer was clearly observed: fluid domains were lower than the crystalline domains. After introducing to the bilayer a very dilute solution of streptavidin to give a final concentration of approximately 0.5 microM, the recognition reaction was imaged in real time. Several hours later, we observed a contrast reversal, i.e., the previously lower fluid domains grew so much in height that they became higher than the crystalline domains. We found that the streptavidin molecules bound almost exclusively to the biotin in the fluid domain (less than 0.25% coverage of the crystalline domains). The apparent structure of the few streptavidin molecules bound to the crystalline domain of the bilayer is shown to depend on the applied force. Finally, in a 2-dimensional quasi-crystal in which the streptavidin molecules were compressed at the air-water interface molecular resolution was achieved.
Weisenhorn
A L
AL
Department of Physics, University of California, Santa Barbara 93106-9530.
Schmitt
F J
FJ
Knoll
W
W
Hansma
P K
PK
D001426:Bacterial Proteins / Q000378:metabolism / Q000648:ultrastructure*; D001710:Biotin; D008051:Lipid Bilayers / Q000378:metabolism; D008853:Microscopy / Q000379:methods; D011485:Protein Binding; D019809:Streptavidin
D001426:Bacterial Proteins; D008051:Lipid Bilayers; D001710:Biotin; D019809:Streptavidin