Generation of longer emission wavelength red fluorescent proteins using computationally designed libraries.

ABSTRACT: The longer emission wavelengths of red fluorescent proteins (RFPs) make them attractive for whole-animal imaging because cells are more transparent to red light. Although several useful RFPs have been developed using directed evolution, the quest for further red-shifted and improved RFPs continues. Herein, we report a structure-based rational design approach to red-shift the fluorescence emission of RFPs. We applied a combined computational and experimental approach that uses computational protein design as an in silico prescreen to generate focused combinatorial libraries of mCherry mutants. The computational procedure helped us identify residues that could fulfill interactions hypothesized to cause red-shifts without destabilizing the protein fold. These interactions include stabilizatio

SUBMITTER: Chica RA 

PROVIDER: S-EPMC2996648 | biostudies-literature | 2010 Nov

REPOSITORIES: biostudies-literature