title: Profiling of bacterial adhesin--host receptor recognition by soluble immunoglobulin superfamily domains.
abstract: Several gram-negative human pathogens recognize members of the carcinoembryonic antigen-related cell adhesion molecule (CEACAM) family. Pathogenic Neisseriae employ distinct isoforms of the colony opacity-associated proteins (Opa(CEA) proteins) to bind to the amino-terminal domains of CEACAMs. Here we present a novel approach to rapidly determine the CEACAM-binding properties of single bacteria. Expression of the isolated amino-terminal domains of various CEACAMs in eukaryotic cells yields soluble probes that selectively recognize Opa(CEA)-expressing bacteria in a pull-down assay format. Furthermore, by expressing soluble CEACAMs as fusions to green-fluorescent protein (CEACAM-N-GFP), CEACAM-binding bacteria can be decorated with a fluorescent label and analysed by flow cytometry allowing the specific detection of receptor binding events on the level of single bacteria. Besides its potential for rapid and quantitative analysis of pathogen-receptor interactions, this novel approach allows the detection of receptor recognition in heterogeneous bacterial populations and might represent a valuable tool for profiling the host binding capabilities of various microorganisms.
authors.affiliation: Lehrstuhl für Zellbiologie, Fachbereich Biologie X908, Universitätsstr.10, Universität Konstanz, 78457 Konstanz, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Kuespert
authors.forename: Stephanie
authors.initials: S
authors.lastname: Weibel
authors.forename: Christof R
authors.initials: CR
authors.lastname: Hauck
mesh_terms: D018829:Adhesins, Bacterial / Q000378:metabolism*; D015703:Antigens, CD / Q000737:chemistry / Q000378:metabolism*; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D005434:Flow Cytometry / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D009344:Neisseria gonorrhoeae / Q000378:metabolism; D009345:Neisseria meningitidis / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D018829:Adhesins, Bacterial; D015703:Antigens, CD; D001425:Bacterial Outer Membrane Proteins; C069847:CD66 antigens; D015815:Cell Adhesion Molecules; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; C090348:Opa protein, Neisseria