title: Relaxed cleavage specificity of an immunoglobulin A1 protease from Neisseria meningitidis.
abstract: Respiratory pathogens, such as Neisseria meningitidis, secrete site-specific proteases able to cleave human immunoglobulin A1 (IgA1), the first line of defense at mucosal membranes. Bacterial isolates show wide variability in IgA1 protease activity, and those isolated from patients with clinical infection possess the highest levels of activity. A feature of this enzyme is the self-cleavage required for secretion of the mature extracellular form. Known cleavage targets contain a proline-rich consensus recognition sequence, Pro-Pro-Ser-Pro, residing in the variable linker region that connects the protease and translocator domains. Here, we report the sequence of the NMB IgA1 protease and the unexpected self-cleavage and subsequent extracellular release of mature IgA1 protease from mutants lacking the previously defined consensus cleavage site. We investigated the possible link between enzyme secretion and variability in the linker sequence segment using site-directed mutagenesis and linker domain swapping to construct mutated and chimeric forms of the IgA1 protease from N. meningitidis strain NMB. The observed change in secreted activity levels compared to the wild-type clone indicated that the precise amino acid sequence of the intervening region, between mature IgA1 protease and the beta-core translocator domain, influences the efficacy of autoproteolytic processing. The broader specificity uncovered for the NMB IgA1 protease suggests that it could cleave a far wider range of human proteins than previously appreciated.
authors.affiliation: Section of Infection, Inflammation and Immunity, Henry Wellcome Laboratories for Medical Research, The University of Sheffield School of Medicine and Biomedical Science, Sheffield, UK.
authors.forename: Srdjan
authors.initials: S
authors.lastname: Vitovski
authors.forename: Jon R
authors.initials: JR
authors.lastname: Sayers
mesh_terms: D003001:Cloning, Molecular; D007070:Immunoglobulin A / Q000235:genetics / Q000378:metabolism*; D009345:Neisseria meningitidis / Q000201:enzymology* / Q000502:physiology; D012697:Serine Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D007070:Immunoglobulin A; D012697:Serine Endopeptidases; C021433:IgA-specific serine endopeptidase