title: Mutation of the zinc-binding metalloprotease motif affects Bacteroides fragilis toxin activity but does not affect propeptide processing.
abstract: To evaluate the role of the zinc-binding metalloprotease in Bacteroides fragilis toxin (BFT) processing and activity, the zinc-binding consensus sequences (H348, E349, H352, G355, H358, and M366) were mutated by site-directed-mutagenesis. Our results indicated that single point mutations in the zinc-binding metalloprotease motif do not affect BFT processing but do reduce or eliminate BFT biologic activity in vitro.
authors.affiliation: Division of Infectious Diseases, Johns Hopkins University School of Medicine, Ross Bldg., Rm. 1167, 1147B Rutland Ave., Baltimore, MD 21205, USA. afranco@jhem.jhmi.edu
authors.forename: Augusto A
authors.initials: AA
authors.lastname: Franco
authors.forename: Simy L
authors.initials: SL
authors.lastname: Buckwold
authors.forename: Jai W
authors.initials: JW
authors.lastname: Shin
authors.forename: Miguel
authors.initials: M
authors.lastname: Ascon
authors.forename: Cynthia L
authors.initials: CL
authors.lastname: Sears
mesh_terms: D020816:Amino Acid Motifs; D001427:Bacterial Toxins / Q000378:metabolism*; D001441:Bacteroides fragilis / Q000201:enzymology; D015820:Cadherins / Q000378:metabolism; D015151:Immunoblotting; D008666:Metalloendopeptidases / Q000378:metabolism*; D045726:Metalloproteases / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D010455:Peptides / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015032:Zinc / Q000378:metabolism*
chemical_list: D001427:Bacterial Toxins; D015820:Cadherins; D010455:Peptides; D045726:Metalloproteases; C405031:Bacteroides fragilis toxin; D008666:Metalloendopeptidases; D015032:Zinc