Iron acquisition systems are critical for bacterial pathogenesis and thus have been proposed as attractive targets for iron-dependent pathogen control. Of these systems, high-affinity iron acquisition mediated by siderophore, a small iron chelator, is the most efficient iron-scavenging mechanism in gram-negative bacteria. Campylobacter does not produce any siderophores but has the ability to utilize exogenous siderophores. In particular, the enterobactin (Ent)-mediated iron scavenging is tightly linked to Campylobacter pathogenesis. To date, Ent, a triscatecholate with the highest known affinity for ferric iron, is a well-characterized siderophore used by Campylobacter for iron acquisition during in vivo infection. Here, we describe the key methods used to characterize Ent-mediated high affinity iron acquisition system in Campylobacter jejuni.
Keywords: Conjugation; Enterobactin; Growth promotion assay; Insertional mutagenesis; Site-directed amino acid substitution mutagenesis.