Green Fluorescent Protein in Inertially Injected Aqueous Nanodroplets
Jianyong Tang, Ana Jofre, G M. Lowman, Rani B. Kishore, Joseph E. Reiner, Kristian Helmerson, Lori S. Goldner, M E. Greene
individual hydrosomes into an immiscible solvent matrix composed of perfluorotriamylamine. Hydrosomes containing EGFP were optically trapped held at the focus of an excitation laser in a confocal microscope, and single molecule photobleaching events were observed. The rotational diffusion time in of EGFP in hydrosomes was measured using time-resolved fluorescence anisotropy. In free solution, the mean rotational correlation time for EGFP at 3 M was measured to be 13.9 0.1 ns. In hydrosomes at the same concentration, the rotational correlation time was measured to be 14.0 0.6 ns. We conclude that the protein does not congregate at the water-solvent interface and that the rotational motion inside the nanodroplets is consistent with free solution. This liberates single molecule protein studies from surface attachment methodologies and opens up new avenues of research using single molecules contained in biologically relevant fluid nanovolumes. Hydrosomes can also be maneuvered and fused in perfluorinated solvents, suggesting their utility for studies of single molecular complex formation and nanochemistry through hydrosome mixing.
Journal of the American Chemical Society
biophysics, fluorescence anisotropy, green fluorescent protein, microemulsion, optical trapping, single molecules
, Jofre, A.
, Lowman, G.
, Kishore, R.
, Reiner, J.
, Helmerson, K.
, Goldner, L.
and Greene, M.
Green Fluorescent Protein in Inertially Injected Aqueous Nanodroplets, Journal of the American Chemical Society
(Accessed May 29, 2023)