ArticleProceedings of the National Academy of Sciences of the United States of America1998
Measurement of cytosolic, mitochondrial, and Golgi pH in single living cells with green fluorescent proteins.
Article in Proceedings of the National Academy of Sciences of the United States of America, 1998. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 385 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
385 citing papers in PubMed, 1,129 citations in OpenAlex.
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- A Color-Tuning Bioluminescent Sensor (AmyLuc) for Real-Time Monitoring of Intracellular pH Dynamics in Cancer Cells.Analytical chemistry · 2026Article
- Reactive sulfur species are inactivated and excreted as trimethylsulfonium ion by thiopurine S-methyltransferase.Redox biology · 2026Article
- Stress-induced proteome remodeling at the Golgi-endosome interface.Journal of cell science · 2026Article
- Designer Aromatic Cations for Photoinduced Protein Ligation, Imaging, and Intracellular Labeling at Extended Wavelengths.Journal of the American Chemical Society · 2026Article
- Stacked-overlapped graphdiyne nano-iontronics enabling enhanced monovalent/divalent cation selectivity for single-cell pH detection.National science review · 2026Article
- DNA nanodevices detect an acidic nanolayer on the lysosomal surface.Nature cell biology · 2026Article
- Organelles harbour pH gradients.Research square · 2026Article
- Lessons in longevity from blood stem cells under protein stress.Trends in cell biology · 2026Review
- A unified intracellular pH landscape with SITE-pHorin: a quantum-entanglement-enhanced pH probe.Science China. Life sciences · 2026Article
- Article
- Organelles harbour pH gradients.bioRxiv : the preprint server for biology · 2025Article
- Nonionotropic action of an acid-sensing ion channel inhibits leukemogenesis in the acidic bone marrow niche.The Journal of clinical investigation · 2025Article
- Article
- Designer Aromatic Cations for Photo-Induced Protein Ligation, Imaging, and Intracellular Labelling at Extended Wavelengths.bioRxiv : the preprint server for biology · 2025Article
- The pH-dependent photophysical and spectral properties of pH-sensing green fluorescent proteins.Physiological reports · 2025Article
- CaBiomolecules · 2025Review
- Exosome-Mediated Mitochondrial Delivery of Antisense Oligonucleotides.Nucleic acid therapeutics · 2025Article
325 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
Many cellular events depend on a tightly compartmentalized distribution of H+ ions across membrane-bound organelles. However, measurements of organelle pH in living cells have been scarce. Several mutants of the Aequorea victoria green fluorescent protein (GFP) displayed a pH-dependent absorbance and fluorescent emission, with apparent pKa values ranging from 6.15 (mutations F64L/S65T/H231L) and 6.4 (K26R/F64L/S65T/Y66W/N146I/M153T/ V163A/N164H/H231L) to a remarkable 7.1 (S65G/S72A/T203Y/H231L). We have targeted these GFPs to the cytosol plus nucleus, the medial/trans-Golgi by fusion with galactosyltransferase, and the mitochondrial matrix by using the targeting signal from subunit IV of cytochrome c oxidase. Cells in culture transfected with these cDNAs displayed the expected subcellular localization by light and electron microscopy and reported local pH that was calibrated in situ with ionophores. We monitored cytosolic and nuclear pH of HeLa cells, and mitochondrial matrix pH in HeLa cells and in rat neonatal cardiomyocytes. The pH of the medial/trans-Golgi was measured at steady-state (calibrated to be 6.58 in HeLa cells) and after various manipulations. These demonstrated that the Golgi membrane in intact cells is relatively permeable to H+, and that Cl- serves as a counter-ion for H+ transport and likely helps to maintain electroneutrality. The amenability to engineer GFPs to specific subcellular locations or tissue targets using gene fusion and transfer techniques should allow us to examine pH at sites previously inaccessible.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.