cb5b00346_si_001pdf. (626.4 kB)
A Fluorescent, Reagentless Biosensor for ATP, Based on Malonyl-Coenzyme A Synthetase
Version 2 2015-12-17, 10:49
Version 1 2015-11-20, 11:35
journal contribution
posted on 2015-12-17, 10:49 authored by Renée Vancraenenbroeck, Martin R. WebbA fluorescent reagentless biosensor
for ATP has been developed,
based on malonyl-coenzyme A synthetase from Rhodopseudomonas
palustris as the protein scaffold and recognition element.
Two 5-iodoacetamidotetramethylrhodamines were covalently bound to
this protein to provide the readout. This adduct couples ATP binding
to a 3.7-fold increase in fluorescence intensity with excitation at
553 nm and emission at 575 nm. It measures ATP concentrations with
micromolar sensitivity and is highly selective for ATP relative to
ADP. Its ability to monitor enzymatic ATP production or depletion
was demonstrated in steady-state kinetic assays in which ATP is a
product or substrate, respectively.