Redox sensor proteins for highly sensitive direct imaging of intracellular redox state

Kazunori Sugiura; Takeharu Nagai; Masahiro Nakano; Hiroshi Ichinose; Takakazu Nakabayashi; Nobuhiro Ohta; Toru Hisabori

doi:10.1016/j.bbrc.2014.12.095

Redox sensor proteins for highly sensitive direct imaging of intracellular redox state

Biochem Biophys Res Commun. 2015 Feb 13;457(3):242-8. doi: 10.1016/j.bbrc.2014.12.095. Epub 2015 Jan 13.

Authors

Kazunori Sugiura¹, Takeharu Nagai², Masahiro Nakano², Hiroshi Ichinose³, Takakazu Nakabayashi⁴, Nobuhiro Ohta⁵, Toru Hisabori⁶

Affiliations

¹ Chemical Resources Laboratory, Tokyo Institute of Technology, Nagatsuta 4259-R1-8, Midori-Ku, Yokohama 226-8503, Japan; Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuta 4259, Midori-Ku, Yokohama, 226-8501, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Tokyo 102-0075, Japan.
² The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka 567-0047, Japan.
³ Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuta 4259, Midori-Ku, Yokohama, 226-8501, Japan.
⁴ Research Institute for Electronic Science, Hokkaido University, Kita 20, Nishi 10, Kita-Ku, Sapporo 001-0020, Japan; Graduate School of Pharmaceutical Sciences, Tohoku University, Aoba-Ku, Sendai 980-8578, Japan.
⁵ Research Institute for Electronic Science, Hokkaido University, Kita 20, Nishi 10, Kita-Ku, Sapporo 001-0020, Japan.
⁶ Chemical Resources Laboratory, Tokyo Institute of Technology, Nagatsuta 4259-R1-8, Midori-Ku, Yokohama 226-8503, Japan; Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuta 4259, Midori-Ku, Yokohama, 226-8501, Japan; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Tokyo 102-0075, Japan. Electronic address: thisabor@res.titech.ac.jp.

PMID: 25592971
DOI: 10.1016/j.bbrc.2014.12.095

Abstract

Intracellular redox state is a critical factor for fundamental cellular functions, including regulation of the activities of various metabolic enzymes as well as ROS production and elimination. Genetically-encoded fluorescent redox sensors, such as roGFP (Hanson, G. T., et al. (2004)) and Redoxfluor (Yano, T., et al. (2010)), have been developed to investigate the redox state of living cells. However, these sensors are not useful in cells that contain, for example, other colored pigments. We therefore intended to obtain simpler redox sensor proteins, and have developed oxidation-sensitive fluorescent proteins called Oba-Q (oxidation balance sensed quenching) proteins. Our sensor proteins derived from CFP and Sirius can be used to monitor the intracellular redox state as their fluorescence is drastically quenched upon oxidation. These blue-shifted spectra of the Oba-Q proteins enable us to monitor various redox states in conjunction with other sensor proteins.

Keywords: Biosensor; Disulfide; Fluorescence; Oxidation-reduction; Redox regulation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Biosensing Techniques / methods
Glutathione / metabolism
Glutathione Disulfide / metabolism
Green Fluorescent Proteins / genetics
Green Fluorescent Proteins / metabolism*
HeLa Cells
Humans
Intracellular Fluid / metabolism*
Mutagenesis
Oxidation-Reduction
Quantum Theory
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / metabolism
Spectrometry, Fluorescence

Substances

Recombinant Fusion Proteins
Green Fluorescent Proteins
Glutathione
Glutathione Disulfide