title: Borrelia burgdorferi binds plasminogen, resulting in enhanced penetration of endothelial monolayers.
abstract: Several strains of Borrelia burgdorferi and Borrelia hermsii can bind human Lys-plasminogen specifically. Affinity blots using 125I-labeled plasminogen showed that numerous polypeptides of all the strains and species tested could bind via lysine residues to the plasminogen molecule since binding could be completely inhibited by the lysine analog epsilon-aminocaproic acid. Binding analysis using 125I-labeled plasminogen on live intact organisms showed that the organisms possess two binding sites for plasminogen: a high-affinity site with a Kd of 24 +/- 12 pM and 106 +/- 14 binding sites per spirochete and a low-affinity site with a Kd of 20 +/- 4 nM and 2,683 +/- 36 binding sites per spirochete. Indirect immunofluorescence and confocal microscopy showed a generalized but punctate pattern of plasminogen binding to the spirochete surface. Exogenously provided urokinase-type plasminogen activator converted B. burgdorferi surface-bound plasminogen to enzymatically active plasmin as demonstrated by the breakdown of the chromogenic plasmin substrate S2251. Plasmin-coated organisms showed an enhanced ability to penetrate endothelial cell monolayers grown on connective tissue substrates compared to untreated controls (P < 0.001). This functional assay demonstrated that enzymatically active plasmin on the surface of spirochetes can lead to greater invasion of tissues.
authors.affiliation: State of New York Department of Health, Stony Brook, USA.
authors.forename: J L
authors.initials: JL
authors.lastname: Coleman
authors.forename: T J
authors.initials: TJ
authors.lastname: Sellati
authors.forename: J E
authors.initials: JE
authors.lastname: Testa
authors.forename: R R
authors.initials: RR
authors.lastname: Kew
authors.forename: M B
authors.initials: MB
authors.lastname: Furie
authors.forename: J L
authors.initials: JL
authors.lastname: Benach
mesh_terms: D000650:Amnion / Q000382:microbiology; D001422:Bacterial Adhesion; D001898:Borrelia / Q000378:metabolism*; D015748:Borrelia burgdorferi Group / Q000378:metabolism*; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000382:microbiology*; D005341:Fibrinolysin / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D010958:Plasminogen / Q000378:metabolism*; D011485:Protein Binding; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism
chemical_list: D010958:Plasminogen; D005341:Fibrinolysin; D014568:Urokinase-Type Plasminogen Activator