8W02

Q108K:K40L:T51V:T53S:R58W mutant of hCRBPII bound to synthetic fluorophore TD-1V

This is a non-PDB format compatible entry.

Summary for 8W02

• Entry DOI: 10.2210/pdb8w02/pdb
• Descriptor: Retinol-binding protein 2, ACETATE ION, (2E)-3-{5-[4-(dimethylamino)phenyl]thiophen-2-yl}but-2-enal, ... (4 entities in total)
• Functional Keywords: human cellular retinol binding protein ii, hcrbpii, td-1v, engineered protein, fluorescent protein, retinol binding protein
• Biological source: Homo sapiens (human)
• Total number of polymer chains: 2
• Total formula weight: 31580.46

Authors

Ghanbarpour, A.,Geiger, J.H.,Borhan, B. (deposition date: 2024-02-13, release date: 2024-04-03, Last modification date: 2024-10-09)

Primary citation

Santos, E.M.,Chandra, I.,Assar, Z.,Sheng, W.,Ghanbarpour, A.,Bingham, C.,Vasileiou, C.,Geiger, J.H.,Borhan, B.
Regulation of Absorption and Emission in a Protein/Fluorophore Complex.
Acs Chem.Biol., 19:1725-1732, 2024

PubMed Abstract: Human cellular retinol binding protein II (hCRBPII) was used as a protein engineering platform to rationally regulate absorptive and emissive properties of a covalently bound fluorogenic dye. We demonstrate the binding of a thio-dapoxyl analog via formation of a protonated imine between an active site lysine residue and the chromophore's aldehyde. Rational manipulation of the electrostatics of the binding pocket results in a 204 nm shift in absorption and a 131 nm shift in emission. The protein is readily expressed in mammalian systems and binds with exogenously delivered fluorophore as demonstrated by live-cell imaging experiments.

PubMed: 39046136
DOI: 10.1021/acschembio.4c00125

Experimental method

X-RAY DIFFRACTION (1.499 Å)