Construction of a thiamin sensor from the periplasmic thiamin binding protein
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This communication describes the development of a thiamin sensor based on the bacterial thiamin binding protein. A triple mutant (C48S, C50S, S62C) of TbpA was labeled on C62 with N-[2-(L-maleimidyl)ethyl]-7-(diethylamino)coumarin-3-carboxamide (MDCC). Thiamin binding to this protein reduced the coumarin fluorescence giving a thiamin sensor with low nanomolar sensitivity.
author list (cited authors)
Hanes, J. W., Chatterjee, D., Soriano, E. V., Ealick, S. E., & Begley, T. P.