Development of an Optical Zn²⁺ Probe Based on a Single Fluorescent Protein.

ABSTRACT: Various fluorescent probes have been developed to reveal the biological functions of intracellular labile Zn²⁺. Here, we present Green Zinc Probe (GZnP), a novel genetically encoded Zn²⁺ sensor design based on a single fluorescent protein (single-FP). The GZnP sensor is generated by attaching two zinc fingers (ZF) of the transcription factor Zap1 (ZF1 and ZF2) to the two ends of a circularly permuted green fluorescent protein (cpGFP). Formation of ZF folds induces interaction between the two ZFs, which induces a change in the cpGFP conformation, leading to an increase in fluorescence. A small sensor library is created to include mutations in the ZFs, cpGFP and linkers between ZF and cpGFP to improve signal stability, sensor brightness and dynamic range based on ration

SUBMITTER: Qin Y 

PROVIDER: S-EPMC5081268 | biostudies-literature | 2016 Oct

REPOSITORIES: biostudies-literature