title: S100 to receptor for advanced glycation end-products binding assay: looking for inhibitors.
abstract: Secreted by tumor and stromal cells, S100 proteins exert their biological functions via the interaction with surface receptors. The most described receptor is the receptor for advanced glycation end-products (RAGE), thereby participating in the S100-dependent cell migration, invasion, tumor growth, angiogenesis and metastasis. Several approaches have been described for determining this interaction. Here we describe an easy, specific and highly reproducible ELISA-based method, by optimizing several parameters such as the binding and blocking buffer, interaction time and concentrations, directed to screen chemical and biological inhibitors of this interaction for S100A4, S100A7 and S100P proteins. The efficiency of the protocol was validated by using well described neutralizing agents of the RAGE receptor and of the S100A4 activity. The methodology described here will allow future works with other members of the S100 protein family and their receptors.
authors.affiliation: Biomed Division of LEITAT Technological Center, Barcelona Science Park, 08028 Barcelona, Spain.
authors.forename: Laura
authors.initials: L
authors.lastname: Padilla
authors.affiliation: Biomed Division of LEITAT Technological Center, Barcelona Science Park, 08028 Barcelona, Spain.
authors.forename: Sheila
authors.initials: S
authors.lastname: Dakhel
authors.affiliation: Biomed Division of LEITAT Technological Center, Barcelona Science Park, 08028 Barcelona, Spain. Electronic address: jlhernandez@leitat.org.
authors.forename: Jose Luis
authors.initials: JL
authors.lastname: Hernández
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D002135:Calcium-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D002450:Cell Communication / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D017127:Glycation End Products, Advanced / Q000378:metabolism; D006801:Humans; D051379:Mice; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D000075023:S100 Calcium Binding Protein A7; D000071999:S100 Calcium-Binding Protein A4; D009418:S100 Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D002135:Calcium-Binding Proteins; D017127:Glycation End Products, Advanced; D009363:Neoplasm Proteins; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic; D011994:Recombinant Proteins; D000075023:S100 Calcium Binding Protein A7; D000071999:S100 Calcium-Binding Protein A4; D009418:S100 Proteins; C089242:S100A7 protein, human; C103003:S100P protein, human; C470944:S100a4 protein, mouse; C077591:S100A4 protein, human
keywords: ELISA; Inhibitors; Interaction; RAGE; S100