title: Identification of a key residue for oligomerisation and pore-formation of Clostridium perfringens NetB.
abstract: Necrotic enteritis toxin B (NetB) is a β-pore-forming toxin produced by Clostridium perfringens and has been identified as a key virulence factor in the pathogenesis of avian necrotic enteritis, a disease causing significant economic damage to the poultry industry worldwide. In this study, site-directed mutagenesis was used to identify amino acids that play a role in NetB oligomerisation and pore-formation. NetB K41H showed significantly reduced toxicity towards LMH cells and human red blood cells relative to wild type toxin. NetB K41H was unable to oligomerise and form pores in liposomes. These findings suggest that NetB K41H could be developed as a genetic toxoid vaccine to protect against necrotic enteritis.
authors.affiliation: College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, UK. s.p.fernandes-da-costa@exeter.ac.uk.
authors.forename: Sérgio P
authors.initials: SP
authors.lastname: Fernandes da Costa
authors.affiliation: Department of Biological Sciences, School of Crystallography, Institute of Structural and Molecular Biology, Birkbeck College, Malet Street, London WC1E 7HX, UK. c.savva@mail.cryst.bbk.ac.uk.
authors.forename: Christos G
authors.initials: CG
authors.lastname: Savva
authors.affiliation: College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, UK. m.bokori-brown@exeter.ac.uk.
authors.forename: Monika
authors.initials: M
authors.lastname: Bokori-Brown
authors.affiliation: Department of Biological Sciences, School of Crystallography, Institute of Structural and Molecular Biology, Birkbeck College, Malet Street, London WC1E 7HX, UK. c.naylor@mail.cryst.bbk.ac.uk.
authors.forename: Claire E
authors.initials: CE
authors.lastname: Naylor
authors.affiliation: Department of Biological Sciences, School of Crystallography, Institute of Structural and Molecular Biology, Birkbeck College, Malet Street, London WC1E 7HX, UK. d.moss@bbk.ac.uk.
authors.forename: David S
authors.initials: DS
authors.lastname: Moss
authors.affiliation: Department of Biological Sciences, School of Crystallography, Institute of Structural and Molecular Biology, Birkbeck College, Malet Street, London WC1E 7HX, UK. a.basak@mail.cryst.bbk.ac.uk.
authors.forename: Ajit K
authors.initials: AK
authors.lastname: Basak
authors.affiliation: College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, UK. r.w.titball@exeter.ac.uk.
authors.forename: Richard W
authors.initials: RW
authors.lastname: Titball
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D001427:Bacterial Toxins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D002645:Chickens; D004768:Enterotoxins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism; D005452:Fluoresceins / Q000378:metabolism; D006461:Hemolysis; D006801:Humans; D052899:Pore Forming Cytotoxic Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017433:Protein Structure, Secondary
chemical_list: D000596:Amino Acids; D001427:Bacterial Toxins; D004768:Enterotoxins; D005452:Fluoresceins; C534774:NetB protein, Clostridium perfringens; D052899:Pore Forming Cytotoxic Proteins; C007740:fluorexon