Structure-guided evolution of Green2 toward photostability and quantum yield enhancement by F145Y substitution.

ABSTRACT: Quantum yield is a determinant for fluorescent protein (FP) applications and enhancing FP brightness through gene engineering is still a challenge. Green2, our de novo FP synthesized by microfluidic picoarray and cloning, has a significantly lower quantum yield than enhanced green FP, though they have high homology and share the same chromophore. To increase its quantum yield, we introduced an F145Y substitution into Green2 based on rational structural analysis. Y145 significantly increased the quantum yield (0.22 vs. 0.18) and improved the photostability (t_1/2 , 73.0 s vs. 46.0 s), but did not affect the excitation and emission spectra. Further structural analysis showed that the F145Y substitution resulted in a significant electrical field change in the immediate environment o

SUBMITTER: Sun T 

PROVIDER: S-EPMC7454421 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature