A new class of binding-protein dependent solute transporter exemplified by the TAXI-GltS system from Bordetella pertussis.

Tripartite ATP-independent periplasmic (TRAP) transporters are widespread in prokaryotes, but absent in eukaryotes, and transport various substrates. TRAP transporters are typically composed of a monomeric substrate binding protein (SBP) and a characteristic transmembrane component. Here, we describe the discovery and characterisation of a TRAP SBP from the TAXI subfamily with a previously unidentified architecture. BP0403 from Bordetella pertussis is a predicted lipoprotein with 3 distinct domains; an α/β globular domain, a helical domain and a C-terminal TAXI SBP domain. Characterisation of full-length BP0403 reveals that it forms a stable dimer, and structural modelling coupled with molecular weight analysis reveals that the interdomain helical region is solely responsible for dimerisation. Differential scanning fluorimetry (DSF) and intrinsic tyrosine fluorescence reveal that BP0403 binds L-glutamate with nanomolar affinity. Unexpectedly, genome context analysis of BP0403 reveals no TRAP membrane component genes; instead, we find co-localisation and translational coupling with gltS, encoding a Na/glutamate symporter. In other bacteria, we identified fused BP0403-GltS homologues, strongly suggesting that this constitutes a completely novel SBP-dependent secondary active transporter. Structural comparisons suggest GltS operates by an elevator-type mechanism, like TRAP transporters; the association of an SBP with this class of secondary transporter is an emerging theme.