Single-molecule dynamics of phytochrome-bound fluorophores probed by fluorescence correlation spectroscopy

Miller AE, Fischer AJ, Laurence T, Hollars CW, Saykally RJ, Lagarias JC, Huser T (2006)
Proceedings of the National Academy of Sciences 103(30): 11136-11141.

Journal Article | Published | English

No fulltext has been uploaded

Author
; ; ; ; ; ;
Abstract
Fluorescence correlation spectroscopy (FCS) was used to investigate the hydrodynamic and photophysical properties of PR1 (phytofluor red 1), an intensely red fluorescent biliprotein variant of the truncated cyanobacterial phytochrome 1 (Cph1 Delta, which consists of the N-terminal 514 amino acids). Single-molecule diffusion measurements showed that PR1 has excellent fluorescence properties at the single-molecule level, making it an interesting candidate for red fluorescent protein fusions. FCS measurements for probing dimer formation in solution over a range of protein concentrations were enabled by addition of Cph1 Delta apoprotein (apoCph1 Delta) to nanomolar solutions of PR1. FCS brightness analysis showed that heterodimerization of PR1 with apoCph1 Delta altered the chemical environment of the PR1 chromophore to further enhance its fluorescence emission. Fluorescence correlation measurements also revealed interactions between apoCph1 Delta and the red fluorescent dyes Cy5.18 and Atto 655 but not Alexa Fluor 660. The concentration dependence of protein:dye complex formation indicated that Atto 655 interacted with, or influenced the formation of, the apoCph1 dimer. These studies presage the utility of phytofluor tags for probing single-molecule dynamics in living cells in which the fluorescence signal can be controlled by the addition of various chromophores that have different structures and photophysical properties, thereby imparting different types of information, such as dimer formation or the presence of open binding faces on a protein.
Publishing Year
ISSN
eISSN
PUB-ID

Cite this

Miller AE, Fischer AJ, Laurence T, et al. Single-molecule dynamics of phytochrome-bound fluorophores probed by fluorescence correlation spectroscopy. Proceedings of the National Academy of Sciences. 2006;103(30):11136-11141.
Miller, A. E., Fischer, A. J., Laurence, T., Hollars, C. W., Saykally, R. J., Lagarias, J. C., & Huser, T. (2006). Single-molecule dynamics of phytochrome-bound fluorophores probed by fluorescence correlation spectroscopy. Proceedings of the National Academy of Sciences, 103(30), 11136-11141.
Miller, A. E., Fischer, A. J., Laurence, T., Hollars, C. W., Saykally, R. J., Lagarias, J. C., and Huser, T. (2006). Single-molecule dynamics of phytochrome-bound fluorophores probed by fluorescence correlation spectroscopy. Proceedings of the National Academy of Sciences 103, 11136-11141.
Miller, A.E., et al., 2006. Single-molecule dynamics of phytochrome-bound fluorophores probed by fluorescence correlation spectroscopy. Proceedings of the National Academy of Sciences, 103(30), p 11136-11141.
A.E. Miller, et al., “Single-molecule dynamics of phytochrome-bound fluorophores probed by fluorescence correlation spectroscopy”, Proceedings of the National Academy of Sciences, vol. 103, 2006, pp. 11136-11141.
Miller, A.E., Fischer, A.J., Laurence, T., Hollars, C.W., Saykally, R.J., Lagarias, J.C., Huser, T.: Single-molecule dynamics of phytochrome-bound fluorophores probed by fluorescence correlation spectroscopy. Proceedings of the National Academy of Sciences. 103, 11136-11141 (2006).
Miller, Abigail E., Fischer, Amanda J., Laurence, Ted, Hollars, Christopher W., Saykally, Richard J., Lagarias, J. Clark, and Huser, Thomas. “Single-molecule dynamics of phytochrome-bound fluorophores probed by fluorescence correlation spectroscopy”. Proceedings of the National Academy of Sciences 103.30 (2006): 11136-11141.
This data publication is cited in the following publications:
This publication cites the following data publications:

12 Citations in Europe PMC

Data provided by Europe PubMed Central.

Fast Photochemistry of Prototypical Phytochromes-A Species vs. Subunit Specific Comparison.
Ihalainen JA, Takala H, Lehtivuori H., Front Mol Biosci 2(), 2015
PMID: 26779488
Removal of Chromophore-Proximal Polar Atoms Decreases Water Content and Increases Fluorescence in a Near Infrared Phytofluor.
Lehtivuori H, Bhattacharya S, Angenent-Mari NM, Satyshur KA, Forest KT., Front Mol Biosci 2(), 2015
PMID: 26636092
Origins of fluorescence in evolved bacteriophytochromes.
Bhattacharya S, Auldridge ME, Lehtivuori H, Ihalainen JA, Forest KT., J. Biol. Chem. 289(46), 2014
PMID: 25253687
Heterogeneous photodynamics of the pfr state in the cyanobacterial phytochrome Cph1.
Kim PW, Rockwell NC, Martin SS, Lagarias JC, Larsen DS., Biochemistry 53(28), 2014
PMID: 24940993
Dynamic inhomogeneity in the photodynamics of cyanobacterial phytochrome Cph1.
Kim PW, Rockwell NC, Martin SS, Lagarias JC, Larsen DS., Biochemistry 53(17), 2014
PMID: 24742290
Single-particle analysis reveals shutoff control of the Arabidopsis ammonium transporter AMT1;3 by clustering and internalization.
Wang Q, Zhao Y, Luo W, Li R, He Q, Fang X, Michele RD, Ast C, von Wiren N, Lin J., Proc. Natl. Acad. Sci. U.S.A. 110(32), 2013
PMID: 23882074
Working memory and decision processes in visual area v4.
Hayden BY, Gallant JL., Front Neurosci 7(), 2013
PMID: 23550043
Characterizing diffusion dynamics of a membrane protein associated with nanolipoproteins using fluorescence correlation spectroscopy.
Gao T, Blanchette CD, He W, Bourguet F, Ly S, Katzen F, Kudlicki WA, Henderson PT, Laurence TA, Huser T, Coleman MA., Protein Sci. 20(2), 2011
PMID: 21280134
Cyanochromes are blue/green light photoreversible photoreceptors defined by a stable double cysteine linkage to a phycoviolobilin-type chromophore.
Ulijasz AT, Cornilescu G, von Stetten D, Cornilescu C, Velazquez Escobar F, Zhang J, Stankey RJ, Rivera M, Hildebrandt P, Vierstra RD., J. Biol. Chem. 284(43), 2009
PMID: 19671704
Nano-biophotonics: new tools for chemical nano-analytics.
Huser T., Curr Opin Chem Biol 12(5), 2008
PMID: 18786651
Characterization of two thermostable cyanobacterial phytochromes reveals global movements in the chromophore-binding domain during photoconversion.
Ulijasz AT, Cornilescu G, von Stetten D, Kaminski S, Mroginski MA, Zhang J, Bhaya D, Hildebrandt P, Vierstra RD., J. Biol. Chem. 283(30), 2008
PMID: 18480055

33 References

Data provided by Europe PubMed Central.

A light-switchable gene promoter system.
Shimizu-Sato S, Huq E, Tepperman JM, Quail PH., Nat. Biotechnol. 20(10), 2002
PMID: 12219076
Synthetic biology: engineering Escherichia coli to see light.
Levskaya A, Chevalier AA, Tabor JJ, Simpson ZB, Lavery LA, Levy M, Davidson EA, Scouras A, Ellington AD, Marcotte EM, Voigt CA., Nature 438(7067), 2005
PMID: 16306980
Fast, flexible algorithm for calculating photon correlations.
Laurence TA, Fore S, Huser T., Opt Lett 31(6), 2006
PMID: 16544638

Rigler R., Mets U., Widengren J., Kask P.., 1993

Widengren J., Mets U., Rigler R.., 1995
Distribution of molecular aggregation by analysis of fluctuation moments.
Qian H, Elson EL., Proc. Natl. Acad. Sci. U.S.A. 87(14), 1990
PMID: 2371284

Export

0 Marked Publications

Open Data PUB

Web of Science

View record in Web of Science®

Sources

PMID: 16844775
PubMed | Europe PMC

Search this title in

Google Scholar