Measuring complexation by single-molecule fluorescence anisotropy.
The complexation of a fluorescent probe by a target protein was observed by single-molecule fluorescence anisotropy. Free and bound states, heterogeneities, and rare binding events can all be observed by this approach. Fluorophore-conjugated biotin was used to bind to NeutrAvidin as a proof-of-concept case. Molecular interactions were observed that could not be elucidated with conventional (ensemble) measurements.
Burrows
Sean M
SM
Department of Chemistry and Biochemistry, Texas Tech University, Box 41061, Lubbock, TX 79409-1061, USA.
Pappas
Dimitri
D
D000818:Animals; D001360:Avidin / Q000737:chemistry; D001710:Biotin / Q000737:chemistry; D005454:Fluorescence Polarization; D006801:Humans; D036103:Nanotechnology; D011485:Protein Binding; D012235:Rhodamines / Q000737:chemistry
D012235:Rhodamines; C093483:neutravidin; D001360:Avidin; D001710:Biotin