A Conserved Mechanism of GABA Binding and Antagonism Is Revealed by Structure-Function Analysis of the Periplasmic Binding Protein Atu2422 in Agrobacterium tumefaciens |
| |
Authors: | Sara Planamente Armelle Vigouroux Samuel Mondy Magali Nicaise Denis Faure Solange Moréra |
| |
Affiliation: | From the ‡Institut des Sciences du Végétal and ;§Laboratoire d''Enzymologie et Biochimie Structurales, CNRS, Avenue de la Terrasse, 91198 Gif-sur-Yvette and ;the ¶Institut de Biochimie et de Biophysique Moléculaire et Cellulaire, UMR8619, CNRS, Université Paris-Sud, 91405 Orsay, France |
| |
Abstract: | Bacterial periplasmic binding proteins (PBPs) and eukaryotic PBP-like domains (also called as Venus flytrap modules) of G-protein-coupled receptors are involved in extracellular GABA perception. We investigated the structural and functional basis of ligand specificity of the PBP Atu2422, which is implicated in virulence and transport of GABA in the plant pathogen Agrobacterium tumefaciens. Five high-resolution x-ray structures of Atu2422 liganded to GABA, Pro, Ala, and Val and of point mutant Atu2422-F77A liganded to Leu were determined. Structural analysis of the ligand-binding site revealed two essential residues, Phe77 and Tyr275, the implication of which in GABA signaling and virulence was confirmed using A. tumefaciens cells expressing corresponding Atu2422 mutants. Phe77 restricts ligand specificity to α-amino acids with a short lateral chain, which act as antagonists of GABA signaling in A. tumefaciens. Tyr275 specifically interacts with the GABA γ-amino group. Conservation of these two key residues in proteins phylogenetically related to Atu2422 brought to light a subfamily of PBPs in which all members could bind GABA and short α-amino acids. This work led to the identification of a fingerprint sequence and structural features for defining PBPs that bind GABA and its competitors and revealed their occurrence among host-interacting proteobacteria. |
| |
Keywords: | Amino Acid Transport Crystal Structure GABA Receptors Plant Transport Amino Acids Host-Pathogen Relationship Periplasmic Binding Protein Quorum Sensing Solute-binding Protein Virulence |
|
|