title: Structural analysis of the Leptospiral sphingomyelinases: in silico and experimental evaluation of Sph2 as an Mg-dependent sphingomyelinase.
abstract: BACKGROUND
  Leptospiral sphingomyelinases are candidate virulence factors present only in pathogenic Leptospira spp. Leptospira interrogans serovar Lai encodes Sph1, Sph2, Sph3, Sph4 and SphH. Except for Sph4, they all possess the exo-endo-phosphatase domain that groups them under the DNase I superfamily.
  
  
  CONCLUSIONS
  Modeling of exo-endo-phosphatase domains reveals high-level structural similarity of Sph2 with the crystal structure of SmcL and BC SMase sphingomyelinases from Listeria ivanovii and Bacillus cereus, respectively. A β-hairpin loop, essential for host cell membrane interaction, is absent in leptospiral sphingomyelinases. Instead, several aromatic amino acids were oriented outward from the surface of these molecules and formed clusters of hydrophobic regions that possibly enables the anchoring of these molecules into the host cell membrane, as demonstrated in Sph2 and Sph3. Sph2 is unique and possesses the Mg(++)-binding Glu53 residue in the metal-binding site and two His residues (His151 and His286) in the catalytic site. We demonstrate experimentally the Mg(++)-dependent hemolysis of erythrocytes by rSph2 and its ability to cleave sphingomyelin to ceramide. Anti-Sph2 antibodies neutralized the hemolytic activity of Sph2. In conclusion, we provide evidence showing that Sph2 is a Mg(++)-dependent hemolysin with both sphingomyelinase and hemolytic activities.
authors.affiliation: Department of Animal Sciences, School of Life Sciences, University of Hyderabad, Hyderabad, India.
authors.forename: Suneel A
authors.initials: SA
authors.lastname: Narayanavari
authors.forename: Nanda M
authors.initials: NM
authors.lastname: Kishore
authors.forename: M
authors.initials: M
authors.lastname: Sritharan
mesh_terms: D001409:Bacillus cereus / Q000201:enzymology; D002518:Ceramides / Q000378:metabolism; D003067:Coenzymes / Q000378:metabolism*; D019295:Computational Biology; D006460:Hemolysin Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006461:Hemolysis; D007921:Leptospira interrogans / Q000201:enzymology* / Q000235:genetics; D008087:Listeria / Q000201:enzymology; D008274:Magnesium / Q000378:metabolism*; D008958:Models, Molecular; D013108:Sphingomyelin Phosphodiesterase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D013109:Sphingomyelins / Q000378:metabolism
chemical_list: D002518:Ceramides; D003067:Coenzymes; D006460:Hemolysin Proteins; D013109:Sphingomyelins; D013108:Sphingomyelin Phosphodiesterase; D008274:Magnesium