Premium
A fluorescent glutathione analog for monitoring interactions of GST fusion proteins
Author(s) -
Huff Hannah C,
Gaines Carson R,
Caldwell Benjamin D
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.613.6
Subject(s) - fusion protein , glutathione , chemistry , biochemistry , glutathione s transferase , fluorescence , fusion , target protein , green fluorescent protein , recombinant dna , biophysics , biology , enzyme , gene , physics , linguistics , philosophy , quantum mechanics
Glutathione S‐Transferase (GST) has commonly been used as a carrier protein in the expression of fusion proteins due to its ease of purification. We are attempting to develop a generic system by which a single probe might be used to monitor many different GST fusion proteins rather than relying on specifically targeted probes which can add expense and time to studies. A probe targeted at GST may reduce the cost and time of screening multiple GST fusion proteins. A fluorescently labeled analog of glutathione was synthesized for monitoring the interaction of GST fusion proteins with target proteins. We will describe the synthesis of fluorescein labeled glutathione (GSH‐Fl) and the purification and characterization of the labeled GST substrate. Initial studies examining the binding of the GSH‐Fl to GST and GST fusions proteins using standard fluorescence and fluorescence anisotropy assays will be described, and its potential to measure protein interactions of GST fusion proteins with target proteins will be discussed.