While the role of A. haemolyticum PLD in pathogenesis is currently unclear, PLD is expressed during infection, as determined by the presence of serum antibodies in pharyngitis patients [15, 16]. PLDs are ubiquitous enzymes which cleave phospholipids, including phosphatidylcholine (PC) and sphingomyelin

(SM), both learn more of which are abundant in the mammalian plasma membrane [17]. SM, with cholesterol and GPI-anchored proteins, predominantly partitions to lipid rafts, which are tightly packed, membrane micro-domains that act to compartmentalize cellular processes on the outer leaflet of the plasma membrane [18]. Lipid rafts are also implicated in host cell invasion by microorganisms [19]. Host PLD cleaves SM releasing ceramide and accumulation of ceramide within

rafts alters their biophysical properties, leading to the formation of large, ceramide-rich membrane platforms [20]. These platforms allow reorganization and aggregation of protein receptors and receptor-associated signaling molecules, which in turn facilitates efficient signal transduction for normal physiological processes [20]. In contrast, PC found in the liquid disordered, or non-raft, phase, is associated with both the inner and outer membrane leaflets, and is cleaved by PLD AZD7762 research buy to phosphatidic acid and choline, which also have roles as second Bioactive Compound Library research buy messengers [18]. PLD is the only A. haemolyticum virulence factor cloned and sequenced to date [21]. Almost invariantly, PLDs possess two His-X-Lys-X4-Asp (HKD) motifs that are involved in catalysis [22]. However, the PLD expressed by A. haemolyticum is not related to these more common HKD PLDs and has a limited substrate specificity which includes SM, but not PC [23], leading to the alternate nomenclature, sphingomyelinase D. Unlike host sphingomyelinases, A. haemolyticum PLD

cleaves SM releasing ceramide-1-PO4 instead of ceramide. Like ceramide, ceramide-1-PO4 is a bioactive sphingolipid, and it acts as a signaling molecule involved in regulating critical cell functions [24]. A. haemolyticum PLD is most closely Glutamate dehydrogenase related to the PLD of Corynebacterium pseudotuberculosis [21]. In C. pseudotuberculosis, PLD is absolutely required for virulence, as a pld mutant could not spread from the site of inoculation or persist in the lymph nodes [25]. C. pseudotuberculosis PLD hydrolyzes SM in host cell membranes and lysophosphatidylcholine in plasma [23], which causes endothelial membrane leakage and cytolysis, leading to enhanced vascular permeability [25]. C. pseudotuberculosis PLD also activates complement [26], promotes neutrophil chemotaxis [27] and is directly dermonecrotic when injected into the skin [26]. The PLDs of recluse spider (Loxosceles spp.) venom are also structurally and functionally related to the A. haemolyticum and corynebacterial PLDs [28].