Water-soluble through-bond energy transfer cassettes for intracellular imaging. Academic Article uri icon

abstract

  • A special, water-soluble, fluorescent probe 1 was designed. This consisted of a fluorescein-based component to harvest irradiation at 488 nm and a rhodamine-based part designed to emit it at a significantly longer wavelength. This cassette was used to label an illustrative protein called ACBP. Evidence was accumulated to support the assertion that ACBP-1 bound its native ligand with a binding constant similar to that of the unlabeled protein, and retained its secondary structure (CD). ACBP-1 was imported into cells using the Chariot peptide. Confocal images proved that some ACBP-1 localized into the nucleus (as expected) and, most significantly, it could be visualized more effectively by irradiating at the donor (fluorescein-like) part of the cassette, than the acceptor (rhodamine-like) part. Overall, this study demonstrates that cassettes of this kind can label a protein without significantly perturbing its function or secondary structure and they can be visualized effectively via irradiation of the donor and observation of the acceptor fluorescence.

published proceedings

  • J Am Chem Soc

author list (cited authors)

  • Bandichhor, R., Petrescu, A. D., Vespa, A., Kier, A. B., Schroeder, F., & Burgess, K.

citation count

  • 111

complete list of authors

  • Bandichhor, Rakeshwar||Petrescu, Anca D||Vespa, Aude||Kier, Ann B||Schroeder, Friedhelm||Burgess, Kevin

publication date

  • August 2006