HyPer (protein)

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GFP from Aequorea victoria , wild-type form.

HyPer is a modified variant of the green fluorescent protein (more precisely: of YFP), in which the fluorescence changes redox-dependently .

properties

HyPer is a fusion protein of cpYFP and the hydrogen peroxide- sensitive bacterial protein OxyR . The oxidation leads to a spectral shift of the absorption of the HyPer from 420 nm to 500 nm.

Applications

The HyPer is used, among other things, as a redox-dependent biosensor and as a reporter protein. With HyPer, the hydrogen peroxide-dependent redox potential can be determined in vivo . In the endoplasmic reticulum , the formation of hydrogen peroxide can be monitored with HyPer. Alternative optical detection methods use e.g. B. Resazurin (synonym AlamarBlue), dichlorofluorescein (DCF) or the proteins roGFP or rxYFP .

Individual evidence

  1. VV Belousov, AF Fradkov, KA Lukyanov, DB Staroverov, KS Shakhbazov, AV Terskikh, S. Lukyanov: Genetically encoded fluorescent indicator for intracellular hydrogen peroxide. In: Nature Methods . Volume 3, Number 4, April 2006, pp. 281-286, ISSN  1548-7091 . doi : 10.1038 / nmeth866 . PMID 16554833 .
  2. KA Lukyanov, VV Belousov: Genetically encoded fluorescent redox sensors. In: Biochimica et Biophysica Acta . Volume 1840, Number 2, February 2014, pp. 745-756, ISSN  0006-3002 . doi : 10.1016 / j.bbagen.2013.05.030 (free full text). PMID 23726987 .
  3. Meyer, AJ, Brach, T., Marty, L., Kreye, S., Rouhier, N., Jacquot, JP, and Hell, R .: Redox-sensitive GFP in Arabidopsis thaliana is a quantitative biosensor for the redox potential of the cellular glutathione redox buffer. . In: Plant J . 52, No. 5, 2007, pp. 973-86. doi : 10.1111 / j.1365-313X.2007.03280.x . PMID 17892447 .
  4. ^ SG Rhee, TS Chang, W. Jeong, D. Kang: Methods for detection and measurement of hydrogen peroxide inside and outside of cells. In: Molecules and cells. Volume 29, Number 6, June 2010, pp. 539-549, ISSN  0219-1032 . doi : 10.1007 / s10059-010-0082-3 . PMID 20526816 .
  5. I. Mehmeti, S. Lortz, S. Lenzen: The H2O2-sensitive HyPer protein targeted to the endoplasmic reticulum as a mirror of the oxidizing thiol-disulfide milieu. In: Free Radical Biology and Medicine . Volume 53, Number 7, October 2012, pp. 1451-1458, ISSN  1873-4596 . doi : 10.1016 / j.freeradbiomed.2012.08.010 . PMID 22921589 .
  6. M. Exposito-Rodriguez, PP Laissue, GR Littlejohn, N. Smirnoff, PM Mullineaux: The use of HyPer to examine spatial and temporal changes in H2O2 in high light-exposed plants. In: Methods in Enzymology . Volume 527, 2013, pp. 185-201, ISSN  1557-7988 . doi : 10.1016 / B978-0-12-405882-8.00010-6 . PMID 23830632 .
  7. S. Pouvreau: Genetically encoded reactive oxygen species (ROS) and redox indicators. In: Biotechnology journal. Volume 9, Number 2, February 2014, pp. 282-293, ISSN  1860-7314 . doi : 10.1002 / biot.201300199 . PMID 24497389 .