compare

Comparison List

mRFP1

a.k.a. mRFP

mRFP1 is a basic (constitutively fluorescent) red fluorescent protein published in 2002, derived from Discosoma sp.. It is reported to be a somewhat slowly-maturing monomer with low acid sensitivity.
+
Aggregation Organism Molecular Weight Cofactor
Monomer Discosoma sp. 25.4 kDa -

Attributes

Ex λ Em λ EC (M-1 cm-1) QY Brightness pKa Maturation (min) Lifetime (ns)
584 607 50,000 0.25 12.5 4.5 60.0  

OSER Measurements

% Normal Cells OSER/NE ratio Cell Type Reference
95.8 ± 1.1 (10000 cells) - HeLa Cranfill et al. (2016)

Photostability

t1/2 (s) Power Light Mode In Cell Fusion ˚C Reference
6.2 Shaner et al. (2004)

mRFP1 Sequence

mRFP1 was derived from dimer1 with the following mutations: V1adel/V71A/K83L/C117E/F124L/V127T/L150M/R153E/V156A/H162K/Q163M/A164R/L174D/V175A/F177V/I180T/Y192A/Y194K/V195T/T197I/H222S/L223T/F224G/L225A
amino acid numbers relative to DsRed. show relative to dimer1

MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMRLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHSTGA
GenBank: CAH64892
IPG: 570910

Excerpts

We present here the stepwise evolution of DsRed to a dimer and then either to a genetic fusion of two copies of the protein, i.e., a tandem dimer, or to a true monomer designated mRFP1 (monomeric red fluorescent protein). Each subunit interface was disrupted by insertion of arginines, which initially crippled the resulting protein, but red fluorescence could be rescued by random and directed mutagenesis totaling 17 substitutions in the dimer and 33 in mRFP1.

Campbell et al. (2002)

Although mRFP1 has somewhat lower extinction coefficient, quantum yield, and photostability than DsRed, mRFP1 matures >10 times faster, so that it shows similar brightness in living cells.

Campbell et al. (2002)

Our basic strategy for decreasing the oligomeric state of DsRed was to replace key dimer interface residues with arginine. When the targeted residue interacts with the identical residue of the dimer partner through symmetry, the high energetic cost of placing two positive charges in close proximity should disrupt the interaction. Such a strategy disrupted the weak tendency of wtGFP to dimerize, without any deleterious effects on GFP maturation or brightness (Zacharias et al. 2002).

Campbell et al. (2002)

Primary Reference

A monomeric red fluorescent protein

Campbell Re, Tour O, Palmer Ae, Steinbach Pa, Baird Gs, Zacharias Da, Tsien Ry

(2002). Proceedings of the National Academy of Sciences, 99(12) , 7877-7882. doi: 10.1073/pnas.082243699. Article   Pubmed

Additional References

  1. Two-photon absorption properties of fluorescent proteins

    Drobizhev M, Makarov Ns, Tillo Se, Hughes Te, Rebane A

    (2011). Nature Methods, 8(5) , 393-399. doi: 10.1038/nmeth.1596. Article   Pubmed

External Resources

Something missing or incorrect? Submit a change Submit a change