Systematic Comparison of Vesicular Targeting Signals Leads to the Development of Genetically Encoded Vesicular Fluorescent Zn²⁺ and pH Sensors.

ABSTRACT: Genetically encoded fluorescent sensors have been widely used to illuminate secretory vesicle dynamics and the vesicular lumen, including Zn²⁺ and pH, in living cells. However, vesicular sensors have a tendency to mislocalize and are susceptible to the acidic intraluminal pH. In this study, we performed a systematic comparison of five different vesicular proteins to target the fluorescent protein mCherry and a Zn²⁺ Förster resonance energy transfer (FRET) sensor to secretory vesicles. We found that motifs derived from vesicular cargo proteins, including chromogranin A (CgA), target vesicular puncta with greater efficacy than transmembrane proteins. To characterize vesicular Zn²⁺ levels, we developed CgA-Zn²⁺ FRET sensor fusions with existing sens

SUBMITTER: Pratt EPS 

PROVIDER: S-EPMC8444068 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature