Generation of bright monomeric red fluorescent proteins via computational design of enhanced chromophore packing.

ABSTRACT: Red fluorescent proteins (RFPs) have found widespread application in chemical and biological research due to their longer emission wavelengths. Here, we use computational protein design to increase the quantum yield and thereby brightness of a dim monomeric RFP (mRojoA, quantum yield = 0.02) by optimizing chromophore packing with aliphatic residues, which we hypothesized would reduce torsional motions causing non-radiative decay. Experimental characterization of the top 10 designed sequences yielded mSandy1 (λ _em = 609 nm, quantum yield = 0.26), a variant with equivalent brightness to mCherry, a widely used RFP. We next used directed evolution to further increase brightness, resulting in mSandy2 (λ _em = 606 nm, quantum yield = 0.35), the brightest Dis

SUBMITTER: Legault S 

PROVIDER: S-EPMC8809391 | biostudies-literature | 2022 Feb

REPOSITORIES: biostudies-literature