Molecular characterization of the interaction of sialic acid with the periplasmic binding protein from Haemophilus ducreyi.


Abstract

The primary role of bacterial periplasmic binding proteins is sequestration of essential metabolites present at a low concentration in the periplasm and making them available for active transporters that transfer these ligands into the bacterial cell. The periplasmic binding proteins (SiaPs) from the tripartite ATP-independent periplasmic (TRAP) transport system that transports mammalian host-derived sialic acids have been well studied from different pathogenic bacteria, including Haemophilus influenzae, Fusobacterium nucleatum, Pasteurella multocida, and Vibrio cholerae SiaPs bind the sialic acid N-acetylneuraminic acid (Neu5Ac) with nanomolar affinity by forming electrostatic and hydrogen-bonding interactions. Here, we report the crystal structure of a periplasmic binding protein (SatA) of the ATP-binding cassette (ABC) transport system from the pathogenic bacterium Haemophilus ducreyi The structure of Hd-SatA in the native form and sialic acid-bound forms (with Neu5Ac and N-glycolylneuraminic acid (Neu5Gc)), determined to 2.2, 1.5, and 2.5 Å resolutions, respectively, revealed a ligand-binding site that is very different from those of the SiaPs of the TRAP transport system. A structural comparison along with thermodynamic studies suggested that similar affinities are achieved in the two classes of proteins through distinct mechanisms, one enthalpically driven and the other entropically driven. In summary, our structural and thermodynamic characterization of Hd-SatA reveals that it binds sialic acids with nanomolar affinity and that this binding is an entropically driven process. This information is important for future structure-based drug design against this pathogen and related bacteria.

Submission Details

ID: DbMXzMbs3

Submitter: Shu-Ching Ou

Submission Date: March 6, 2019, 5:33 p.m.

Version: 1

Publication Details
Gangi Setty T;Mowers JC;Hobbs AG;Maiya SP;Syed S;Munson RS Jr;Apicella MA;Subramanian R,J Biol Chem (2018) Molecular characterization of the interaction of sialic acid with the periplasmic binding protein from Haemophilus ducreyi. PMID:30315109
Additional Information

NB means no binding.

Sequence Assay Result Units