Photoactivatable fluorescent protein

Photoactivatable fluorescent proteins (PAFPs) is a type of fluorescent protein that exhibit fluorescence that can be modified by a light-induced chemical reaction.

History

The first PAFP, Kaede (protein), was isolated from Trachyphyllia geoffroyi in a cDNA library screen designed to identify new fluorescent proteins.^[1] A fluorescent green protein derived from this screen was serendipitously discovered to have sensitivity to ultraviolet light--

We happened to leave one of the protein aliquots on the laboratory bench overnight. The next day, we found that the protein sample on the bench had turned red, whereas the others that were kept in a paper box remained green. Although the sky had been partly cloudy, the red sample had been exposed to sunlight through the south-facing windows.^[1]

Properties

Many PAFPs have been engineered from existing fluorescent proteins or identified from large-scale screens in the wake of Kaede's discovery. Many of these undergo green-to-red photoconversion, but other colors are available. Some proteins take part in irreversible photoconversion reactions while other reactions can be reversed using light of a specific wavelength.

List of PAFPs

PAFP Properties^[2]
PAFP	Absorbance₁ (nm)	Emission₁ (nm)	Absorbance₂ (nm)	Emission₂ (nm)	Photoconversion wavelength	Reversibility	Brightness₁*	Brightness₂*	Reference
Kaede (protein)	508	518	572	580	ultraviolet	none	2.64X	0.60X	^[1]
Eos (protein)	506	516	571	581	ultraviolet	none	1.30X	0.70X	^[3]
IrisFP	488	516	551	580	ultraviolet	none	0.66X	0.49X	^[4]
IrisFP	488	516	390	?	490 nm	reversible, 390 nm	?	?	idem
IrisFP	551	580	440	?	550 nm	reversible, 440 nm	?	?	idem
KikGR/Kikume	507	517	583	593	ultraviolet	none	0.60X	0.64X	^[5]
Dronpa	503	518	390	?	490 nm	reversible, 390 nm	?	?	^[6]
PAGFP	400	?	504	517	ultraviolet	none	0.08X	0.42X	^[7]
PS-CFP	402	468	490	511	ultraviolet	none	0.17X	0.16X	^[8]
KFP1	?	?	590	600	green	variable	0.004X	0.13X
*Brightness values are relative to EGFP.

Applications

Unlike other fluorescent proteins, PAFPs can be used as selective optical markers. An entirely labeled cell can be followed to assess cell division, migration, and morphology. Very small volumes containing PAFPs can be activated with a laser. In these cases, protein trafficking, diffusion, and turnover can be assessed.

References

^ ^a ^b ^c Ando, Ryoko; Hama, Hiroshi; Yamamoto-Hino, Miki; Mizuno, Hideaki; Miyawaki, Atsushi (2002-10-01). "An optical marker based on the UV-induced green-to-red photoconversion of a fluorescent protein". Proceedings of the National Academy of Sciences. 99 (20): 12651–12656. Bibcode:2002PNAS...9912651A. doi:10.1073/pnas.202320599. ISSN 0027-8424. PMC 130515. PMID 12271129.
^ Lukyanov, Konstantin A.; Chudakov, Dmitry M.; Lukyanov, Sergey; Verkhusha, Vladislav V. (November 2005). "Photoactivatable fluorescent proteins". Nature Reviews Molecular Cell Biology. 6 (11): 885–890. doi:10.1038/nrm1741. ISSN 1471-0072. PMID 16167053. S2CID 23734846.
^ Wiedenmann, J.; Ivanchenko, S.; Oswald, F.; Schmitt, F.; Rocker, C.; Salih, A.; Spindler, K.-D.; Nienhaus, G. U. (2004-11-09). "EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion". Proceedings of the National Academy of Sciences. 101 (45): 15905–15910. Bibcode:2004PNAS..10115905W. doi:10.1073/pnas.0403668101. ISSN 0027-8424. PMC 528746. PMID 15505211.
^ Adam, Virgile; Lelimousin, Mickaël; Boehme, Susan; Desfonds, Guillaume; Nienhaus, Karin; Field, Martin J.; Wiedenmann, Joerg; McSweeney, Sean; Nienhaus, G. Ulrich; Bourgeois, Dominique (2008-11-25). "Structural characterization of IrisFP, an optical highlighter undergoing multiple photo-induced transformations". Proceedings of the National Academy of Sciences. 105 (47): 18343–18348. Bibcode:2008PNAS..10518343A. doi:10.1073/pnas.0805949105. ISSN 0027-8424. PMC 2587625. PMID 19017808.
^ Tsutsui, Hidekazu; Karasawa, Satoshi; Shimizu, Hideaki; Nukina, Nobuyuki; Miyawaki, Atsushi (March 2005). "Semi-rational engineering of a coral fluorescent protein into an efficient highlighter". EMBO Reports. 6 (3): 233–238. doi:10.1038/sj.embor.7400361. ISSN 1469-221X. PMC 1299271. PMID 15731765.
^ Habuchi, Satoshi; Ando, Ryoko; Dedecker, Peter; Verheijen, Wendy; Mizuno, Hideaki; Miyawaki, Atsushi; Hofkens, Johan (2005-07-05). "Reversible single-molecule photoswitching in the GFP-like fluorescent protein Dronpa". Proceedings of the National Academy of Sciences. 102 (27): 9511–9516. Bibcode:2005PNAS..102.9511H. doi:10.1073/pnas.0500489102. ISSN 0027-8424. PMC 1157093. PMID 15972810.
^ Patterson, George H; Lippincott-Schwartz, Jennifer (April 2004). "Selective photolabeling of proteins using photoactivatable GFP". Methods. 32 (4): 445–450. doi:10.1016/j.ymeth.2003.10.006. PMID 15003607.
^ Chudakov, Dmitriy M; Verkhusha, Vladislav V; Staroverov, Dmitry B; Souslova, Ekaterina A; Lukyanov, Sergey; Lukyanov, Konstantin A (2004-11-01). "Photoswitchable cyan fluorescent protein for protein tracking". Nature Biotechnology. 22 (11): 1435–1439. doi:10.1038/nbt1025. ISSN 1087-0156. PMID 15502815. S2CID 9597405.

[:0-1] Ando, Ryoko; Hama, Hiroshi; Yamamoto-Hino, Miki; Mizuno, Hideaki; Miyawaki, Atsushi (2002-10-01). "An optical marker based on the UV-induced green-to-red photoconversion of a fluorescent protein". Proceedings of the National Academy of Sciences. 99 (20): 12651–12656. Bibcode:2002PNAS...9912651A. doi:10.1073/pnas.202320599. ISSN 0027-8424. PMC 130515. PMID 12271129.

[2] Lukyanov, Konstantin A.; Chudakov, Dmitry M.; Lukyanov, Sergey; Verkhusha, Vladislav V. (November 2005). "Photoactivatable fluorescent proteins". Nature Reviews Molecular Cell Biology. 6 (11): 885–890. doi:10.1038/nrm1741. ISSN 1471-0072. PMID 16167053. S2CID 23734846.

[3] Wiedenmann, J.; Ivanchenko, S.; Oswald, F.; Schmitt, F.; Rocker, C.; Salih, A.; Spindler, K.-D.; Nienhaus, G. U. (2004-11-09). "EosFP, a fluorescent marker protein with UV-inducible green-to-red fluorescence conversion". Proceedings of the National Academy of Sciences. 101 (45): 15905–15910. Bibcode:2004PNAS..10115905W. doi:10.1073/pnas.0403668101. ISSN 0027-8424. PMC 528746. PMID 15505211.

[4] Adam, Virgile; Lelimousin, Mickaël; Boehme, Susan; Desfonds, Guillaume; Nienhaus, Karin; Field, Martin J.; Wiedenmann, Joerg; McSweeney, Sean; Nienhaus, G. Ulrich; Bourgeois, Dominique (2008-11-25). "Structural characterization of IrisFP, an optical highlighter undergoing multiple photo-induced transformations". Proceedings of the National Academy of Sciences. 105 (47): 18343–18348. Bibcode:2008PNAS..10518343A. doi:10.1073/pnas.0805949105. ISSN 0027-8424. PMC 2587625. PMID 19017808.

[5] Tsutsui, Hidekazu; Karasawa, Satoshi; Shimizu, Hideaki; Nukina, Nobuyuki; Miyawaki, Atsushi (March 2005). "Semi-rational engineering of a coral fluorescent protein into an efficient highlighter". EMBO Reports. 6 (3): 233–238. doi:10.1038/sj.embor.7400361. ISSN 1469-221X. PMC 1299271. PMID 15731765.

[6] Habuchi, Satoshi; Ando, Ryoko; Dedecker, Peter; Verheijen, Wendy; Mizuno, Hideaki; Miyawaki, Atsushi; Hofkens, Johan (2005-07-05). "Reversible single-molecule photoswitching in the GFP-like fluorescent protein Dronpa". Proceedings of the National Academy of Sciences. 102 (27): 9511–9516. Bibcode:2005PNAS..102.9511H. doi:10.1073/pnas.0500489102. ISSN 0027-8424. PMC 1157093. PMID 15972810.

[7] Patterson, George H; Lippincott-Schwartz, Jennifer (April 2004). "Selective photolabeling of proteins using photoactivatable GFP". Methods. 32 (4): 445–450. doi:10.1016/j.ymeth.2003.10.006. PMID 15003607.

[8] Chudakov, Dmitriy M; Verkhusha, Vladislav V; Staroverov, Dmitry B; Souslova, Ekaterina A; Lukyanov, Sergey; Lukyanov, Konstantin A (2004-11-01). "Photoswitchable cyan fluorescent protein for protein tracking". Nature Biotechnology. 22 (11): 1435–1439. doi:10.1038/nbt1025. ISSN 1087-0156. PMID 15502815. S2CID 9597405.

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