Bacterial sialidases cleave terminal sialic acid from a variety of host glycoproteins, and contribute to survival and growth of many human-dwelling bacterial species, including various pathogens.Tannerella forsythia, an oral, Gram-negative, fastidious anaerobe, is a key organism in periodontal disease, and possesses a dedicated sialic acid utilisation and scavenging (nan) operon, including NanH sialidase. Here, we describe biochemical characterisation of recombinant NanH, including its action on host-relevant sialoglycans such as sialyl Lewis A and sialyl Lewis X (SLeA/X), and on human cell-attached sialic acids directly, uncovering that it is a highly active broad specificity sialidase. Furthermore, theN-terminal domain of NanH was hypothesised and proven to be capable of binding to a range of sialoglycans and non-sialylated derivatives withKdin the micromolar range, as determined by steady-state tryptophan fluorescence spectroscopy, but it has no catalytic activity in isolation from the active site. We consider this domain to represent the founding member of a novel subfamily of Carbohydrate Binding Module (CBM), involved in glycosidase-ligand binding. In addition, we created a catalytically inactive version of the NanH enzyme (FRIP→YMAP) that retained its ability to bind sialic acid-containing ligands and revealed for the first time that binding activity of a CBM is enhanced by association with the catalytic domain. Finally, we investigated the importance of Lewis-type sialoglycans onT. forsythia-host interactions, showing that nanomolar amounts of SLeA/Xwere capable of reducing invasion of oral epithelial cells byT. forsythiasuggesting that these are key ligands for bacterial-cellular interactions during periodontal disease.
CORE (COnnecting REpositories)
CORE (COnnecting REpositories)