title: Localization of the cleavage site specificity determinant of Haemophilus influenzae immunoglobulin A1 protease genes.
abstract: Immunoglobulin A1 (IgA1) proteases are produced by a number of different species of bacteria which cause infection at human mucosal surfaces. The sole substrate of these proteases is human IgA1. Cleavage is within the hinge region of IgA1, although there is variability in the exact peptide bond within the hinge region that is cut by a particular protease. The cleavage site of the Haemophilus influenzae type 1 protease is located four amino acids from the cleavage site of the type 2 enzyme. In this study, the region of the H. influenzae IgA1 protease gene (iga) that determines the cleavage site specificity was localized through the comparison of the type 1 and type 2 genes and the construction and analysis of type 1-type 2 hybrid genes. The hybrid genes were generated by in vivo and in vitro techniques which facilitated the selection and screening of randomly generated hybrids. The cleavage site determinant was found to be within a 370-base-pair region near the amino-terminal coding region, in one of two large areas of nonhomology between the two types of H. influenzae iga genes. DNA sequence analysis of the cleavage site determinant and surrounding regions did not reveal a simple mechanism whereby one enzyme type could be converted to the other type. Comparison of the type 2 gonococcal IgA1 protease gene to the two Haemophilus genes revealed a significant amount of homology around the cleavage site determinant, with the two type 2 genes showing greater homology.
authors.affiliation: Department of Molecular Biology and Microbiology, Tufts University Health Sciences Campus, Boston, Massachusetts 02111.
authors.forename: F J
authors.initials: FJ
authors.lastname: Grundy
authors.forename: A G
authors.initials: AG
authors.lastname: Plaut
authors.forename: A
authors.initials: A
authors.lastname: Wright
mesh_terms: D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D004269:DNA, Bacterial / Q000032:analysis; D005798:Genes, Bacterial; D006193:Haemophilus influenzae / Q000201:enzymology* / Q000235:genetics; D009344:Neisseria gonorrhoeae / Q000201:enzymology; D009693:Nucleic Acid Hybridization; D010447:Peptide Hydrolases / Q000235:genetics*; D011817:Rabbits; D011995:Recombination, Genetic; D012689:Sequence Homology, Nucleic Acid; D012697:Serine Endopeptidases; D014169:Transformation, Bacterial
chemical_list: D004269:DNA, Bacterial; D010447:Peptide Hydrolases; D012697:Serine Endopeptidases; C021433:IgA-specific serine endopeptidase