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Cited by 408

The DOI matches its Crossref record; 30 of the 30 references it cites were found in a registry or an index.

Imaging Intracellular pH in Live Cells with a Genetically Encoded Red Fluorescent Protein Sensor

Open access
Tantama, Mathew, Hung, Yin Pun, Yellen, Gary·2011·Journal of the American Chemical Society

Abstract

Intracellular pH affects protein structure and function, and proton gradients underlie the function of organelles such as lysosomes and mitochondria. We engineered a genetically encoded pH sensor by mutagenesis of the red fluorescent protein mKeima, providing a new tool to image intracellular pH in live cells. This sensor, named pHRed, is the first ratiometric, single-protein red fluorescent sensor of pH. Fluorescence emission of pHRed peaks at 610 nm while exhibiting dual excitation peaks at 440 and 585 nm that can be used for ratiometric imaging. The intensity ratio responds with an apparent pK(a) of 6.6 and a >10-fold dynamic range. Furthermore, pHRed has a pH-responsive fluorescence lifetime that changes by ~0.4 ns over physiological pH values and can be monitored with single-wavelength two-photon excitation. After characterizing the sensor, we tested pHRed's ability to monitor intracellular pH by imaging energy-dependent changes in cytosolic and mitochondrial pH.

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Metadata

Authors
Tantama, M., Hung, Y. P., & Yellen, G.
Venue
Journal of the American Chemical Society
Published
2011-06-09
Volume
133
Issue
26
Pages
10034-10037
ISSN
  • 0002-7863 (ISSN-L)
  • 1520-5126
OpenAlex
W2027631918
Cited by
408
References
30
FWCI
13.88
Type
journal article
Language
en
Open access
green

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