Universal fluorescent sensors of high-affinity iron transport, applied to ESKAPE pathogens.

ABSTRACT: Sensitive assays of biochemical specificity, affinity, and capacity are valuable both for basic research and drug discovery. We created fluorescent sensors that monitor high-affinity binding reactions and used them to study iron acquisition by ESKAPE bacteria, which are frequently responsible for antibiotic-resistant infections. By introducing site-directed Cys residues in bacterial iron transporters and modifying them with maleimide fluorophores, we generated living cells or purified proteins that bind but do not transport target compounds. These constructs sensitively detected ligand concentrations in solution, enabling accurate, real-time spectroscopic analysis of membrane transport by other cells. We assessed the efficacy of these "fluorescent decoy" (FD) sensors by characterizing acti

SUBMITTER: Chakravorty S 

PROVIDER: S-EPMC6433069 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature