Rational Design of the β-Bulge Gate in a Green Fluorescent Protein Accelerates the Kinetics of Sulfate Sensing.

Autor: Ong WSY; Department of Chemistry and Biochemistry, The University of Texas at Dallas, 800 W. Campbell Road, Richardson, TX 75080-3021, USA., Ji K; Department of Chemistry and Biochemistry, The University of Texas at Dallas, 800 W. Campbell Road, Richardson, TX 75080-3021, USA., Pathiranage V; Department of Chemistry, Wayne State University, 42 W. Warren Ave., Detroit, MI 48202, USA., Maydew C; Department of Chemistry and Biochemistry, The University of Texas at Dallas, 800 W. Campbell Road, Richardson, TX 75080-3021, USA., Baek K; Department of Chemistry and Biochemistry, The University of Texas at Dallas, 800 W. Campbell Road, Richardson, TX 75080-3021, USA., Villones RLE; Department of Chemistry and Biochemistry, The University of Texas at Dallas, 800 W. Campbell Road, Richardson, TX 75080-3021, USA., Meloni G; Department of Chemistry and Biochemistry, The University of Texas at Dallas, 800 W. Campbell Road, Richardson, TX 75080-3021, USA., Walker AR; Department of Chemistry, Wayne State University, 42 W. Warren Ave., Detroit, MI 48202, USA., Dodani SC; Department of Chemistry and Biochemistry, The University of Texas at Dallas, 800 W. Campbell Road, Richardson, TX 75080-3021, USA.
Jazyk: angličtina
Zdroj: Angewandte Chemie (International ed. in English) [Angew Chem Int Ed Engl] 2023 Jun 26; Vol. 62 (26), pp. e202302304. Date of Electronic Publication: 2023 May 16.
DOI: 10.1002/anie.202302304
Abstrakt: Detection of anions in complex aqueous media is a fundamental challenge with practical utility that can be addressed by supramolecular chemistry. Biomolecular hosts such as proteins can be used and adapted as an alternative to synthetic hosts. Here, we report how the mutagenesis of the β-bulge residues (D137 and W138) in mNeonGreen, a bright, monomeric fluorescent protein, unlocks and tunes the anion preference at physiological pH for sulfate, resulting in the turn-off sensor SulfOFF-1. This unprecedented sensing arises from an enhancement in the kinetics of binding, largely driven by position 138. In line with these data, molecular dynamics (MD) simulations capture how the coordinated entry and gating of sulfate into the β-barrel is eliminated upon mutagenesis to facilitate binding and fluorescence quenching.
(© 2023 Wiley-VCH GmbH.)
Databáze: MEDLINE
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