Journal article
The pH-dependent photophysical and spectral properties of pH-sensing green fluorescent proteins
Physiological reports, Vol.13(20), e70625
10/2025
DOI: 10.14814/phy2.70625
PMCID: PMC12535197
PMID: 41108577
Appears in UI Libraries Support Open Access
Abstract
Protonation and deprotonation reactions significantly influence biological function, with buffers and transporters maintaining stable pH levels in the cytosol and various organs. However, pH can vary considerably in organ lumens, such as the airway surface liquid (ASL), which serves as a defense barrier for airways. Traditional microelectrodes, while precise, face limitations prompting the use of fluorescent pH sensors. Although many green fluorescent protein (GFP)-based pH sensors have been reported, data are usually limited to their spectral properties. Photophysical properties are often reported for a single pH value. In this study, the photophysical properties of pHluorins and superfolder GFP-including a novel pH sensor pHluorin4-were compared. Purified pHluorin4 effectively reported ASL pH differences in cystic fibrosis (CF) and non-CF epithelia. These findings provide a comprehensive summary of the photophysical and spectral properties of popular pHluorins, including the new member pHluorin4, which can be used to optimize their application in different systems.
Details
- Title: Subtitle
- The pH-dependent photophysical and spectral properties of pH-sensing green fluorescent proteins
- Creators
- Ian M Thornell - University of Iowa
- Resource Type
- Journal article
- Publication Details
- Physiological reports, Vol.13(20), e70625
- DOI
- 10.14814/phy2.70625
- PMID
- 41108577
- PMCID
- PMC12535197
- NLM abbreviation
- Physiol Rep
- ISSN
- 2051-817X
- eISSN
- 2051-817X
- Publisher
- Wiley
- Grant note
- National Institute of Health: HL007638, HL091842, HL152960 Division of Pulmonary, Critical Care, and Occupational Medicine, Internal Medicine (University of Iowa)Cystic Fibrosis Foundation (Iowa CFF Research Development Program)
IMT was supported, in part, by the National Institute of Health (HL007638) and additional funding by the Division of Pulmonary, Critical Care, and Occupational Medicine, Internal Medicine (University of Iowa). The In Vitro Models Cell Culture Core was supported, in part, by the National Institute of Health (HL091842, HL152960) and by the Cystic Fibrosis Foundation (Iowa CFF Research Development Program).
- Language
- English
- Date published
- 10/2025
- Academic Unit
- Pulmonary, Critical Care, and Occupational Medicine; Internal Medicine
- Record Identifier
- 9985014800502771
Metrics
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