Synthesis of a new water-soluble rhodamine derivative and application to protein labeling and intracellular imaging.
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Synthesis of a new fluorescent rhodamine derivative, dye 1, is reported. This probe is different from other rhodamines insofar as it has several (four) carboxylic acid functionalities to promote water solubility and facilitate conjugation to proteins. It also has an aryl bromide functionality that could, in principle, be used to further functionalize the system for specialized applications. Dye 1 was conjugated to a model protein called ACBP (acyl-CoA binding protein). The properties of this conjugate were tested to establish that the label does not significantly perturb the binding function of the protein to its natural ligand in vitro and to confirm that its secondary structure was not significantly perturbed (circular dichroism). Experiments were performed to test if the labeled protein could be imported into living COS-7 cells (using the Chariot-peptide delivery system) and, if so, to observe, via fluorescence microscopy, which of the labeled protein was able to migrate to the nucleus, as expected for ACBP in cells. In the event, all these postulates were confirmed.
author list (cited authors)
Bandichhor, R., Petrescu, A. D., Vespa, A., Kier, A. B., Schroeder, F., & Burgess, K.
complete list of authors
Bandichhor, Rakeshwar||Petrescu, Anca D||Vespa, Aude||Kier, Ann B||Schroeder, Friedhelm||Burgess, Kevin