In vivo indicators of cytoplasmic, vacuolar, and extracellular pH using pHluorin2 in Candida albicans

Hélène Tournu, Arturo Luna-Tapia, Brian Peters, Glen Palmer

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Environmental or chemically induced stresses often trigger physiological responses that regulate intracellular pH. As such, the capacity to detect pH changes in real time and within live cells is of fundamental importance to essentially all aspects of biology. In this respect, pHluorin, a pH-sensitive variant of green fluorescent protein, has provided an invaluable tool to detect such responses. Here, we report the adaptation of pHluorin2 (PHL2), a substantially brighter variant of pHluorin, for use with the human fungal pathogen Candida albicans. As well as a cytoplasmic PHL2 indicator, we describe a version that specifically localizes within the fungal vacuole, an acidified subcellular compartment with important functions in nutrient storage and pH homeostasis. In addition, by means of a glycophosphatidylinositolanchored PHL2-fusion protein, we generated a cell surface pH sensor. We demonstrated the utility of these tools in several applications, including accurate intracellular and extracellular pH measurements in individual cells via flow cytometry and in cell populations via a convenient plate reader-based protocol. The PHL2 tools can also be used for endpoint as well as time course experiments and to conduct chemical screens to identify drugs that alter normal pH homeostasis. These tools enable observation of the highly dynamic intracellular pH shifts that occur throughout the fungal growth cycle, as well as in response to various chemical treatments.

Original languageEnglish (US)
Article numbere00276-17
JournalmSphere
Volume2
Issue number4
DOIs
StatePublished - Jul 1 2017

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Candida albicans
Homeostasis
Vacuoles
Green Fluorescent Proteins
Flow Cytometry
Observation
Food
Growth
Pharmaceutical Preparations
Population
Proteins

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology

Cite this

In vivo indicators of cytoplasmic, vacuolar, and extracellular pH using pHluorin2 in Candida albicans. / Tournu, Hélène; Luna-Tapia, Arturo; Peters, Brian; Palmer, Glen.

In: mSphere, Vol. 2, No. 4, e00276-17, 01.07.2017.

Research output: Contribution to journalArticle

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