title: Specific binding of human fibrinogen fragment D to Aspergillus fumigatus conidia.
abstract: The interaction of purified human fibrinogen with Aspergillus fumigatus conidia was investigated by immunofluorescence and electron microscopy and binding assays with radiolabeled proteins. We described the localization of the binding sites on the A. fumigatus conidia and on the fibrinogen molecule and determined the binding characteristics. Immunofluorescence revealed that the fixation of purified fibrinogen was selectively associated with conidia and suggested a role for the D domains of the fibrinogen molecule. Binding assays performed with 125I-radiolabeled proteins confirmed that binding sites were located specifically in the D domains. No reaction could be detected with fragment E. The binding of 125I-fragment D to conidia was time dependent, saturable, and specific. Scatchard analysis of the data revealed an average of 1,200 binding sites per conidium, and an apparent dissociation constant (Kd) of 2.2 x 10(-9) M was estimated. Pretreatment of the cells with proteolytic enzymes or heat abolished binding, demonstrating the protein nature of the binding sites. Ultrastructural localization of the fungal receptors was determined by transmission electron microscopy. Labeling appeared to be associated with the outer electron-dense layer of the conidial wall and progressively decreased during the germination process. Labeling of thin sections with fragment D and an antifibrinogen immune serum revealed that binding sites also lay in the inner part of the wall and in vacuoles. These results indicate the presence at the conidial surface of specific receptors for fibrinogen which could act as mediators of conidial adherence to host tissues.
authors.affiliation: Laboratoire de Parasitologie-Mycologie, Centre Hospitalier Régional et Universitaire, Angers, France.
authors.forename: V
authors.initials: V
authors.lastname: Annaix
authors.forename: J P
authors.initials: JP
authors.lastname: Bouchara
authors.forename: G
authors.initials: G
authors.lastname: Larcher
authors.forename: D
authors.initials: D
authors.lastname: Chabasse
authors.forename: G
authors.initials: G
authors.lastname: Tronchin
mesh_terms: D001232:Aspergillus fumigatus / Q000378:metabolism*; D001665:Binding Sites; D005338:Fibrin Fibrinogen Degradation Products / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006801:Humans; D008854:Microscopy, Electron; D010980:Platelet Membrane Glycoproteins / Q000032:analysis*
chemical_list: D005338:Fibrin Fibrinogen Degradation Products; D010980:Platelet Membrane Glycoproteins; C015214:fibrinogen D fragment