Generation and characterization of a lysosomally targeted, genetically encoded Ca<SUP>2+</SUP>-sensor

McCue, Hannah V, Wardyn, Joanna D, Burgoyne, Robert D ORCID: 0000-0002-9219-0387 and Haynes, Lee P ORCID: 0000-0002-1296-0338 (2013) Generation and characterization of a lysosomally targeted, genetically encoded Ca<SUP>2+</SUP>-sensor. BIOCHEMICAL JOURNAL, 449 (2). pp. 449-457.

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Official URL: http://dx.doi.org/10.1042/bj20120898

Abstract

Distinct spatiotemporal Ca2+ signalling events regulate fundamental aspects of eukaryotic cell physiology. Complex Ca2+ signals can be driven by release of Ca2+ from intracellular organelles that sequester Ca2+ such as the ER (endoplasmic reticulum) or through the opening of Ca2+-permeable channels in the plasma membrane and influx of extracellular Ca2+. Late endocytic pathway compartments including late-endosomes and lysosomes have recently been observed to sequester Ca2+ to levels comparable with those found within the ER lumen. These organelles harbour ligand-gated Ca2+-release channels and evidence indicates that they can operate as Ca2+-signalling platforms. Lysosomes sequester Ca2+ to a greater extent than any other endocytic compartment, and signalling from this organelle has been postulated to provide 'trigger' release events that can subsequently elicit more extensive Ca2+ signals from stores including the ER. In order to investigate lysosomal-specific Ca2+ signalling a simple method for measuring lysosomal Ca2+ release is essential. In the present study we describe the generation and characterization of a genetically encoded, lysosomally targeted, cameleon sensor which is capable of registering specific Ca2+ release in response to extracellular agonists and intracellular second messengers. This probe represents a novel tool that will permit detailed investigations examining the impact of lysosomal Ca2+ handling on cellular physiology.

Item Type:	Article
Additional Information:	## TULIP Type: Articles/Papers (Journal) ##
Uncontrolled Keywords:	cameleon, calcium, fluorescence resonance energy transfer (FRET), lysosome, sensor, signalling, targeting
Depositing User:	Symplectic Admin
Date Deposited:	28 Apr 2016 10:18
Last Modified:	07 Oct 2023 16:24
DOI:	10.1042/BJ20120898
Open Access URL:	http://www.ncbi.nlm.nih.gov/pmc/articles/PMC352611...
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3000898