A pH-correctable, DNA-based fluorescent reporter for organellar calcium

It is extremely challenging to quantitate lumenal Ca²⁺ in acidic Ca²⁺ stores of the cell because all Ca²⁺ indicators are pH sensitive, and Ca²⁺ transport is coupled to pH in acidic organelles. We have developed a fluorescent DNA-based reporter, CalipHluor, that is targetable to specific organelles. By ratiometrically reporting lumenal pH and Ca²⁺ simultaneously, CalipHluor functions as a pH-correctable Ca²⁺ reporter. By targeting CalipHluor to the endolysosomal pathway, we mapped lumenal Ca²⁺ changes during endosomal maturation and found a surge in lumenal Ca²⁺ specifically in lysosomes. Using lysosomal proteomics and genetic analysis, we found that catp-6, a Caenorhabditis elegans homolog of ATP13A2, was responsible for lysosomal Ca²⁺ accumulation-an example of a lysosome-specific Ca²⁺ importer in animals. By enabling the facile quantification of compartmentalized Ca²⁺, CalipHluor can expand the understanding of subcellular Ca²⁺ importers.