title: Purification of the proteinase from group B streptococci that inactivates human C5a.
abstract: We previously reported that group B streptococci (GBS) possess a cell-associated activity that inactivates the chemotactic activity generated in zymosan-activated serum by cleaving a specific site within the carboxy termini of C5a and C5adesarg. This inactivates the major chemoattractants for neutrophils that are generated when serum complement is activated. We now report the isolation of the enzyme responsible for the proteolytic cleavage of C5a. Treatment of GBS with mutanolysin, an endo-N-acetyl muramidase, released activity from GBS which destroyed the functional activity of C5a. The soluble activity was purified to homogeneity by hydroxyapatite, ion-exchange and gel-filtration chromatography. Analysis by SDS-PAGE showed that the enzyme (GBS C5a-ase) has an Mr of approx. 120,000. The GBS C5a-ase appears to be a serine esterase on the basis of its sensitivity to di-isopropyl fluorophosphate. This enzyme is distinct from the C5a-cleaving enzyme produced by group A streptococci, since the two bacterial products migrate differently on SDS-PAGE, and lack antigenic cross reactivity. This enzyme may play a role in the pathogenesis of group B streptococcal disease through its ability to rapidly inactivate the potent neutrophil agonist, C5a, at sites of infection.
authors.affiliation: Department of Pediatrics, University of Utah School of Medicine, Salt Lake City 84132.
authors.forename: J F
authors.initials: JF
authors.lastname: Bohnsack
authors.forename: X N
authors.initials: XN
authors.lastname: Zhou
authors.forename: P A
authors.initials: PA
authors.lastname: Williams
authors.forename: P P
authors.initials: PP
authors.lastname: Cleary
authors.forename: C J
authors.initials: CJ
authors.lastname: Parker
authors.forename: H R
authors.initials: HR
authors.lastname: Hill
mesh_terms: D018829:Adhesins, Bacterial; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology; D015936:Complement C5a / Q000037:antagonists & inhibitors*; D010450:Endopeptidases / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology; D006801:Humans; D015151:Immunoblotting; D013292:Streptococcus agalactiae / Q000201:enzymology*
chemical_list: D018829:Adhesins, Bacterial; D015936:Complement C5a; D010450:Endopeptidases; C053088:C5a peptidase; C021816:mutanolysin