Computational redesign of a fluorogen activating protein with Rosetta
Explore the source record for details and available documents.
Bozhanova, Nina G.↗
Engineering topics
Mishin, Alexander S.
Publications and source records attributed to Mishin, Alexander S..
Structure-Based Rational Design of Two Enhanced Bacterial Lipocalin Blc Tags for Protein-PAINT Super
Explore the source record for details and available documents.
Muslinkina, Liya↗
DiB-splits: nature-guided design of a novel fluorescent labeling split system
Fluorogen-activating proteins (FAPs) are innovative fluorescent probes combining advantages of genetically-encoded proteins such as green fluorescent protein and externally added fluorogens that allow for highly tunable and on demand fluorescent signaling. Previously, a panel of green- and red-emitting FAPs has been created from bacterial lipocalin Blc (named DiBs). Here we present a rational design as well as functional and structural characterization of the first self-assembling FAP split system, DiB-splits. This new system decreases the size of the FAP label to ~8–12 kDa while preserving DiBs’ unique properties: strong increase in fluorescence intensity of the chromophore upon binding, binding affinities to the chromophore in nanomolar to low micromolar range, and high photostability of the protein-ligand complex. These properties allow for use of DiB-splits for wide-field, confocal, and super-resolution fluorescence microscopy. DiB-splits also represent an attractive starting point for further design of a protein-protein interaction detection system as well as novel FAP-based sensors.
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗