Here, we use an Escherichia coliΔnanT strain to characterize

the function of known and proposed bacterial sialic acid transporters. We discover that the STM1128 gene from Salmonella enterica serovar Typhimurium, which encodes a member of the sodium solute symporter family, is able to restore growth on sialic acid to the ΔnanT strain and is Epacadostat able to transport [14C]-sialic acid. Using the ΔnanT genetic background, we performed a direct in vivo comparison of the transport properties of the STM1128 protein with those of sialic acid transporters of the major facilitator superfamily and tripartite ATP-independent periplasmic families, E. coli NanT and Haemophilus influenzae SiaPQM, respectively. This revealed that both STM1128 and SiaPQM are sodium-dependent and, unlike SiaPQM, both STM1128 and NanT are reversible secondary carriers, demonstrating qualitative functional differences in the properties of sialic acid transporters

used by bacteria that colonize humans. Sialic acids are a family of related nine carbon sugar acids that play important roles in the biology Tofacitinib ic50 of a wide range of both eukaryotic and prokaryotic organisms (Schauer, 2004; Vimr et al., 2004). In mammals, sialic acids are a predominant feature on the surface of many cell types, and bacteria have evolved multiple mechanisms to exploit these host-derived sugars (Vimr et al., 2004; Severi et al., 2007). For example, Escherichia coli is able to grow on the most common sialic acid N-acetylneuraminic

acid (Neu5Ac) as a sole carbon and nitrogen source (Vimr & Troy, 1985), which is important for successful colonization of the mouse gut (Chang et al., 2004). Other bacteria such as Haemophilus influenzae use host-derived Neu5Ac in an immune evasion mechanism by adding it as a terminal component of their lipopolysaccharide (Bouchet et al., 2003). While some pathogens have evolved de Elongation factor 2 kinase novo biosynthesis pathways for Neu5Ac (Vimr et al., 2004), many bacteria rely on the acquisition of Neu5Ac from their environment and hence require high-affinity transport systems (Bouchet et al., 2003). The pioneering work of Vimr and colleagues led to the first molecular characterization of a bacterial Neu5Ac transporter, which was the NanT protein from E. coli (Vimr & Troy, 1985). This is a secondary transporter and a member of the major facilitator superfamily (MFS) (Martinez et al., 1995). Very recently, another MFS family member, distinct from NanT, has been implicated in sialic acid uptake in Bacteriodes fragilis (Brigham et al., 2009) and Tannerella forsythia (Thompson et al., 2009). We and others have characterized a tripartite ATP-independent periplasmic (TRAP) transporter for Neu5Ac from H. influenzae, SiaPQM, that is important for virulence (Allen et al., 2005; Severi et al., 2005; Mulligan et al., 2009).