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Comparison List

pcDronpa2

pcDronpa2 is a multi-photochromic green fluorescent protein published in 2014, derived from Echinophyllia sp. SC22. It has moderate acid sensitivity.

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Oligomerization Organism Molecular Weight Cofactor
Tetramer Echinophyllia sp. SC22 25.3 kDa -

FPbase ID: GGUJF

Attributes

State Ex λ Em λ EC (M-1 cm-1) QY Brightness pKa Maturation (min) Lifetime (ns)
Green 504 515 100,000 0.83 83.0 5.8    
Green (Off)              
Red 569 583 105,000 0.68 71.4 6.1    

Transitions

From To Switch λ
Green Red 405
Green (Off) Green 405
Green Green (Off) 488

Photostability

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pcDronpa2 Sequence

pcDronpa2 was derived from pcDronpa with the following mutations: Y116N

MSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKVKEGGPLPFAYDILTTAFHYGNRVFAKYPENIVDYFKQSFPEGYSWERSMSYEDGGICIATNDITLDGDCYINEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALSLEGGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSGLPRQAK

Excerpts

The red form of pcDronpa2 has the highest extinction coefficient of all PCFP red forms known to date and is, just like its ancestor pcDronpa, not photoswitchable in the red form. Rational mutagenesis based on literature examples (S142A, V157I/G/S, M159A/T, F173S/L)15,23,45,48 did not generate a four-way highlighter pcDronpa2 mutant (data not shown).

Moeyaert et al. (2014)

We found that reverting the N102I and E218G mutations is not a viable strategy to obtain a monomeric pcDronpa2 variant... As an alternative approach, we used our crystallographic data to rationally break the tetramer interfaces in a different way. Mutations N158E and Y188A disrupted the A/C interface, while V123T additionally broke the A/B interface. Although we could thus successfully make a monomeric version of pcDronpa2 (pcDronpa2-V123T-N158E-Y188A), this mutant displayed no photoconversion.

Moeyaert et al. (2014)

Primary Reference

Additional References

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External Resources

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