title: Separable domains define target cell specificities of an RTX hemolysin from Actinobacillus pleuropneumoniae.
abstract: The leukotoxin (LktA) from Pasteurella haemolytica and the hemolysin (AppA) from Actinobacillus pleuropneumoniae are members of a highly conserved family of cytolytic proteins produced by gram-negative bacteria. Despite the extensive homology between these gene products, LktA is specific for ruminant leukocytes while AppA, like other hemolysins, lyses erythrocytes and a variety of nucleated cells, including ruminant leukocytes. Both proteins require activation facilitated by the product of an accessory repeat toxin (RTX) C gene for optimal biological activity. We have constructed six genes encoding hybrid toxins by recombining domains of ltkA and appA and have examined the target cell specificities of the resulting hybrid proteins. Our results indicate that the leukocytic potential of AppA, like that of LktA, maps to the C-terminal half of the protein and is physically separable from the region specifying erythrocyte lysis. As a consequence, we were able to construct an RTX toxin capable of lysing erythrocytes but not leukocytes. The specificity of one hybrid was found to be dependent upon the RTX C gene used for activation. With appC activation, this hybrid toxin lysed both erythrocytes and leukocytes, while lktC activation produced a toxin which could attack only leukocytes. This is the first demonstration that the specificity of an RTX toxin can be determined by the process of C-mediated activation.
authors.affiliation: Department of Medical Biochemistry and Genetics, College of Medicine, Texas A&M University, College Station 77843.
authors.forename: D R
authors.initials: DR
authors.lastname: McWhinney
authors.forename: Y F
authors.initials: YF
authors.lastname: Chang
authors.forename: R
authors.initials: R
authors.lastname: Young
authors.forename: D K
authors.initials: DK
authors.lastname: Struck
mesh_terms: D016977:Actinobacillus pleuropneumoniae / Q000235:genetics / Q000378:metabolism*; D000371:Agglutination; D001711:Biotransformation; D003603:Cytotoxins / Q000235:genetics / Q000378:metabolism*; D005098:Exotoxins / Q000235:genetics / Q000378:metabolism; D005796:Genes; D005809:Genes, Regulator; D006460:Hemolysin Proteins / Q000235:genetics / Q000378:metabolism; D016978:Mannheimia haemolytica / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D013329:Structure-Activity Relationship
chemical_list: D003603:Cytotoxins; D005098:Exotoxins; D006460:Hemolysin Proteins; D011993:Recombinant Fusion Proteins; C018579:leukotoxin