Photoactivatable fluorescent protein

Photoactivatable fluorescent proteins (PAFPs) 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.[2]

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 photoconversions, 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[3]
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 [4]
Eos (protein) 506 516 571 581 ultraviolet none 1.30X 0.70X [5]
IrisFP 488 516 551 580 ultraviolet none 0.66X 0.49X [6]
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 [7]
Dronpa 503 518 390 ? 490 nm reversible, 390 nm ? ? [8]
PAGFP 400 ? 504 517 ultraviolet none 0.08X 0.42X [9]
PS-CFP 402 468 490 511 ultraviolet none 0.17X 0.16X [10]
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. ↑ Ando et al. 2002 PNAS 99(20) pp 12651-6.
  2. ↑ Ando et al. 2002 PNAS 99(20) p 12652
  3. ↑ Lukyanov et al. 2005. Nature Reviews: Molecular Cell Biology 6(11) pp 885-91.
  4. ↑ Ando et al. 2002 PNAS 99(20) pp 12651-6.
  5. ↑ Wiedenmann et al. 2004. PNAS 101(45), pp 15905-10.
  6. ↑ Adam et al. 2008. PNAS 105(47), pp 18343–48.
  7. ↑ Tsutsui et al. 2005. EMBO Reports 6(3), pp 233-8.
  8. ↑ Habuchi et al. 2005. PNAS 102(27), pp 9511-6.
  9. ↑ Patterson & Lippincott-Schwartz 2004. Methods 32(4), pp 445-50.
  10. ↑ Chudakov et al. 2004. Nature Biotechnology 22(11), pp 1435-9.
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