A xanthene-BODIPY cassette is used as a ratiometric intracellular pH reporter
for imaging protein-dye conjugates in living cells. A model was hypothesized to explain
the pH-dependent energy transfer efficiencies from the donor to the acceptor based on
the electronic chemistry data.
Sulfonation conditions were developed for BODIPY dyes to give water-soluble
functionalized monosulfonation and disulfonation donors. A water-soluble TBET
cassette, which has good photophysical properties, was synthesized using a bissulfonated
BODIPY dye as the donor, and their applications for in vitro protein labeling is
achieved. Chemoselective cross-coupling reactions were demonstrated for C-S bonds in
the BODIPY dye, and similar reactions were applied to make the acceptor of the watersoluble
cassette.
Chemiluminescent energy transfer cassettes based on fluorescein and Nile Red
were synthesized and their spectral properties were studied.
Pep-1 (also known as Chariot), R8 (which is not often used as a non-covalent
protein carrier), and a new synthesized compound, Azo-R8, was used for the study of non-covalent delivery of four different proteins into mammalian cells. Data from
confocal spectroscopy revealed that all three carriers are effective for translocating
protein cargos into live cells. At 37 dgrees C, import into endocytic compartments dominates,
but at 4 degrees C weak, diffuse fluorescence is observed in the cytosol indicative of a favorable
mode of action.
| Identifer | oai:union.ndltd.org:tamu.edu/oai:repository.tamu.edu:1969.1/ETD-TAMU-2009-08-7016 |
| Date | 2009 August 1900 |
| Creators | Han, Junyan |
| Source Sets | Texas A and M University |
| Language | en_US |
| Detected Language | English |
| Type | thesis, text |
| Format | application/pdf |
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