title: Binding and degradation of elastin by the staphylolytic enzyme lysostaphin.
abstract: Lysostaphin is a bacterial zinc metalloproteinase that degrades staphylococcal cell wall peptidoglycans. We have shown that lysostaphin also binds tightly to elastin and contains elastolytic activity. The objective of this investigation was to further characterize the biochemical mechanism of elastolysis by lysostaphin. Binding of lysostaphin to elastin was demonstrated with elastin peptide affinity chromatography. Elastolysis by lysostaphin was studied using a tritium release assay with tritium borohydride-reduced elastin as substrate. Proteolysis of elastin by lysostaphin was maximal at neutral to slightly basic pH and proceeded linearly for 10 hr before reaching saturation. The elastolytic activity was not affected by inhibitors of cysteine or serine proteinases, but was inhibited by o-phenanthroline, EDTA, and by the addition of exogenous zinc. Inhibition by zinc was not due to an alteration in enzyme-ligand interaction since zinc-containing buffers did not impair the ability of lysostaphin to bind elastin. Lysostaphin elastolysis was not inhibited by trypsin treatment of the enzyme, which has been shown to inactivate the enzyme's staphylolytic properties. It is therefore concluded that: (1) lysostaphin binds and degrades elastin; (2) the elastolytic activity of lysostaphin is distinct from its staphylolytic activity; (3) sequence similarities with matrix metalloproteinases suggest the Ala-Ala-Thr-His-Glu sequence in the amino terminus of lysostaphin to be involved in elastin degradation; (4) our studies are important in establishing that metalloproteinases from staphylococci can participate in elastin degradation.
authors.affiliation: Department of Cell Biology and Physiology, Washington University School of Medicine, St Louis, MO 63110, USA.
authors.forename: P W
authors.initials: PW
authors.lastname: Park
authors.forename: R M
authors.initials: RM
authors.lastname: Senior
authors.forename: G L
authors.initials: GL
authors.lastname: Griffin
authors.forename: T J
authors.initials: TJ
authors.lastname: Broekelmann
authors.forename: M S
authors.initials: MS
authors.lastname: Mudd
authors.forename: R P
authors.initials: RP
authors.lastname: Mecham
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002846:Chromatography, Affinity; D004549:Elastin / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008022:Ligaments; D008248:Lysostaphin / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D011485:Protein Binding; D013211:Staphylococcus aureus / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D010446:Peptide Fragments; D004549:Elastin; D008248:Lysostaphin