title: Development of a ligand blot assay using biotinylated live cells.
abstract: Adhesive interactions between cells are critical to a variety of processes, including host-pathogen relationships. The authors have developed a new technique for the observation of binding interactions in which molecules obtained from excised tissues are resolved by gel electrophoresis and transferred to a membrane. Biotinylated live cells are then kept in contact with that membrane, and their interactions with proteins of interest are detected by peroxidase-labeled streptavidin, followed by a biotin-streptavidin detection system. The adhesion proteins can eventually be identified by cutting the relevant band(s) and performing mass spectrometry or other amino acid-sequencing methods. The technique described here allows for the identification of both known and novel adhesion molecules capable of binding to live cells, among a complex mixture and without previous isolation or purification. This is especially important for the analysis of host-parasite interactions and may be extended to other types of cell-cell interactions.
authors.affiliation: Instituto de Microbiologia Prof. Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Felipe
authors.initials: F
authors.lastname: De Almeida Dias
authors.forename: André Luis
authors.initials: AL
authors.lastname: Souza Dos Santos
authors.forename: Antônio
authors.initials: A
authors.lastname: Ferreira-Pereira
authors.forename: Alexandre
authors.initials: A
authors.lastname: Romeiro
authors.forename: Luciana
authors.initials: L
authors.lastname: Teixeira Zimmermann
authors.forename: Michelle Tanny
authors.initials: MT
authors.lastname: Cunha do Nascimento
authors.forename: Georgia
authors.initials: G
authors.lastname: Correa Atella
authors.forename: Elvira Maria
authors.initials: EM
authors.lastname: Saraiva
authors.forename: Rafael
authors.initials: R
authors.lastname: Linden
authors.forename: Angela
authors.initials: A
authors.lastname: Hampshire Lopes
mesh_terms: D000818:Animals; D001710:Biotin / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006790:Host-Parasite Interactions; D007313:Insecta; D008024:Ligands
chemical_list: D008024:Ligands; D001710:Biotin