Auto-luminescent genetically-encoded ratiometric indicator for real-time Ca2+ imaging at the single cell level.

ABSTRACT:

Background

Efficient bioluminescence resonance energy transfer (BRET) from a bioluminescent protein to a fluorescent protein with high fluorescent quantum yield has been utilized to enhance luminescence intensity, allowing single-cell imaging in near real time without external light illumination.

Methodology/principal findings

We applied BRET to develop an autoluminescent Ca(2+) indicator, BRAC, which is composed of Ca(2+)-binding protein, calmodulin, and its target peptide, M13, sandwiched between a yellow fluorescent protein variant, Venus, and an enhanced Renilla luciferase, RLuc8. Adjusting the relative dipole orientation of the luminescent protein's chromophores improved the dynamic range of BRET signal change in BRAC up to 60%, which is the largest dynamic range among

SUBMITTER: Saito K 

PROVIDER: S-EPMC2848576 | biostudies-literature | 2010 Apr

REPOSITORIES: biostudies-literature