Biology: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
- ↑ 1.0 1.1 1.2 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" (in en). Proceedings of the National Academy of Sciences 99 (20): 12651–12656. doi:10.1073/pnas.202320599. ISSN 0027-8424. PMID 12271129. Bibcode: 2002PNAS...9912651A.
- ↑ Lukyanov, Konstantin A.; Chudakov, Dmitry M.; Lukyanov, Sergey; Verkhusha, Vladislav V. (November 2005). "Photoactivatable fluorescent proteins" (in en). Nature Reviews Molecular Cell Biology 6 (11): 885–890. doi:10.1038/nrm1741. ISSN 1471-0072. PMID 16167053. http://www.nature.com/articles/nrm1741.
- ↑ 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" (in en). Proceedings of the National Academy of Sciences 101 (45): 15905–15910. doi:10.1073/pnas.0403668101. ISSN 0027-8424. PMID 15505211. Bibcode: 2004PNAS..10115905W.
- ↑ Adam, Virgile; Lelimousin, Mickaël; Boehme, Susan; Desfonds, Guillaume; Nienhaus, Karin; Field, Martin J.; Wiedenmann, Joerg; McSweeney, Sean et al. (2008-11-25). "Structural characterization of IrisFP, an optical highlighter undergoing multiple photo-induced transformations" (in en). Proceedings of the National Academy of Sciences 105 (47): 18343–18348. doi:10.1073/pnas.0805949105. ISSN 0027-8424. PMID 19017808. Bibcode: 2008PNAS..10518343A.
- ↑ Tsutsui, Hidekazu; Karasawa, Satoshi; Shimizu, Hideaki; Nukina, Nobuyuki; Miyawaki, Atsushi (March 2005). "Semi‐rational engineering of a coral fluorescent protein into an efficient highlighter" (in en). EMBO Reports 6 (3): 233–238. doi:10.1038/sj.embor.7400361. ISSN 1469-221X. 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" (in en). Proceedings of the National Academy of Sciences 102 (27): 9511–9516. doi:10.1073/pnas.0500489102. ISSN 0027-8424. PMID 15972810. Bibcode: 2005PNAS..102.9511H.
- ↑ Patterson, George H; Lippincott-Schwartz, Jennifer (April 2004). "Selective photolabeling of proteins using photoactivatable GFP" (in en). Methods 32 (4): 445–450. doi:10.1016/j.ymeth.2003.10.006. PMID 15003607. https://linkinghub.elsevier.com/retrieve/pii/S1046202303002743.
- ↑ 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" (in en). Nature Biotechnology 22 (11): 1435–1439. doi:10.1038/nbt1025. ISSN 1087-0156. PMID 15502815. http://www.nature.com/articles/nbt1025.
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