Effects of point mutations in the binding pocket of the mouse major urinary protein MUP20 on ligand affinity and specificity

April 8, 2019

Title

Effects of point mutations in the binding pocket of the mouse major urinary protein MUP20 on ligand affinity and specificity

Author

Jimena Ricatti, Laura Acquasaliente, Giovanni Ribaudo, Vincenzo De Filippis, Marino Bellini, Ramiro Esteban Llovera, Susi Barollo, Raffaele Pezzani, Giuseppe Zagotto, Krishna C. Persaud & Carla Mucignat-Caretta

Year

2019

Journal

Scientific Reports

Abstract

The mouse Major Urinary Proteins (MUPs) contain a conserved β-barrel structure with a characteristic central hydrophobic pocket that binds a variety of volatile compounds. After release of urine, these molecules are slowly emitted in the environment where they play an important role in chemical communication. MUPs are highly polymorphic and conformationally stable. They may be of interest in the construction of biosensor arrays capable of detection of a broad range of analytes. In this work, 14 critical amino acids in the binding pocket involved in ligand interactions were identified in MUP20 using in silico techniques and 7 MUP20 mutants were synthesised and characterised to produce a set of proteins with diverse ligand binding profiles to structurally different ligands. A single amino acid substitution in the binding pocket can dramatically change the MUPs binding affinity and ligand specificity. These results have great potential for the design of new biosensor and gas-sensor recognition elements.

Instrument

V-630, J-810

Keywords

Biomimetic synthesis, Proteomics