Journal
NATURE METHODS
Volume 16, Issue 1, Pages 95-+Publisher
NATURE PORTFOLIO
DOI: 10.1038/s41592-018-0232-7
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Funding
- University of Chicago Women's Board
- NIDDK center [P30DK42086]
- MRSEC [DMR-1420709]
- National Center for Advancing Translational Sciences of the National Institutes of Health [1UL1TR002389-01]
- Searle Funds at The Chicago Community Trust [C-084]
- University of Chicago
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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.
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