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Fluorescence Properties of the Chromophore-Binding Domain of Bacteriophytochrome from Deinococcus radiodurans
- Source :
- Journal of Physical Chemistry B, 117(38), 11049-11057. American Chemical Society, Lehtivuori, H, Rissanen, I, Takala, H, Bamford, J K H, Tkachenko, N V & Ihalainen, J A 2013, ' Fluorescence properties of the chromophore-binding domain of bacteriophytochrome from Deinococcus radiodurans ', Journal of Physical Chemistry B, vol. 117, no. 38, pp. 11049-11057 . https://doi.org/10.1021/jp312061b
- Publication Year :
- 2013
- Publisher :
- American Chemical Society (ACS), 2013.
-
Abstract
- Fluorescent proteins are versatile tools for molecular imaging. In this study, we report a detailed analysis of the absorption and fluorescence properties of the chromophore-binding domain from Deinococcus radiodurans and its D207H mutant. Using single photon counting and transient absorption techniques, the average excited state lifetime of both studied systems was about 370 ps. The D207H mutation slightly changed the excited state decay profile but did not have a considerable effect on the average decay time of the system or the shape of the absorption and emission spectra of the biliverdin chromophore. We confirmed that the fluorescence properties of both samples are very similar in vivo and in vitro. However, we found that the paraformaldehyde fixation of the Escherichia coli cells containing the recombinant phytochrome protein significantly changed the fluorescence properties of the chromophore-binding domain. The biliverdin fluorescence was diminished almost completely, and the fluorescence originated only from the protoporphyrin molecules. Our results emphasize that the effect of protoporphyrin IXa should not be ignored in the fluorescence experiments with phytochrome systems while designing better red fluorescence markers for cellular imaging.
- Subjects :
- Time Factors
Fluorescence in the life sciences
Photochemistry
chemistry.chemical_compound
Bimolecular fluorescence complementation
Bacterial Proteins
Escherichia coli
Materials Chemistry
Physical and Theoretical Chemistry
ta116
Biliverdin
biology
Phytochrome
Biliverdine
ta1182
Deinococcus radiodurans
Chromophore
biology.organism_classification
Fluorescence
Recombinant Proteins
Protein Structure, Tertiary
Surfaces, Coatings and Films
chemistry
Mutation
Quantum Theory
Spectrophotometry, Ultraviolet
Deinococcus
Binding domain
Subjects
Details
- ISSN :
- 15205207 and 15206106
- Volume :
- 117
- Database :
- OpenAIRE
- Journal :
- The Journal of Physical Chemistry B
- Accession number :
- edsair.doi.dedup.....ec5091d88cc05388d79b6f04ba17276c