- Construction of a thiamin sensor from the periplasmic thiamin binding protein.
Construction of a thiamin sensor from the periplasmic thiamin binding protein.
Chemical communications (Cambridge, England) (2010-12-03)
Jeremiah W Hanes, Debashree Chatterjee, Erika V Soriano, Steven E Ealick, Tadhg P Begley
PMID21125117
RESUMO
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.
MATERIAIS
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Sigma-Aldrich
7-Diethylamino-3-[N-(2-maleimidoethyl)carbamoyl]coumarin, suitable for fluorescence, BioReagent, ≥97.0% (HPLC)