Development of a range of fluorescent reagentless biosensors for ATP, based on malonyl-coenzyme A synthetase.

ABSTRACT: The range of ATP concentrations that can be measured with a fluorescent reagentless biosensor for ATP has been increased by modulating its affinity for this analyte. The ATP biosensor is an adduct of two tetramethylrhodamines with MatB from Rhodopseudomonas palustris. Mutations were introduced into the binding site to modify ATP binding affinity, while aiming to maintain the concomitant fluorescence signal. Using this signal, the effect of mutations in different parts of the binding site was measured. This mutational analysis revealed three variants in particular, each with a single mutation in the phosphate-binding loop, which had potentially beneficial changes in ATP binding properties but preserving a fluorescence change of ~3-fold on ATP binding. Two variants (T167A and T303A) weakened

SUBMITTER: Vancraenenbroeck R 

PROVIDER: S-EPMC5479551 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature