Title
Stabilization of structure in near-infrared fluorescent proteins by binding of biliverdin chromophore
Author
Olesya V. Stepanenko, Olga V. Stepanenko, G.S. Bublikova, I.M. Kuznetsova, V.V. Verkhusha, K.K. Turoverov
Year
2016
Journal
Journal of Molecular Structure
Abstract
Near-infrared fluorescent proteins (NIR FPs) engineered from bacterial phytochromes and their mutants with different location of Cys residues, which able to bind a biliverdin chromophore, or without these Cys residues were studied using intrinsic tryptophan fluorescence, NIR fluorescence and circular dichroism. It was shown that a covalent binding of the biliverdin chromophore to a Cys residue via thioether group substantially stabilizes the spatial structure of NIR FPs. The stability of the protein structure and the chromophore association strength strongly depends on the location of Cys residues and decreases in the following order: a protein with Cys residues in both domains, a protein with Cys in PAS domains, and a protein with Cys in GAF domains. NIR FPs without Cys residues capable to covalently attach biliverdin have the lowest stability, comparable to NIR FP apoforms.
Instrument
J-810
Keywords
Circular dichroism, Secondary structure, Tertiary structure, Biochemistry