title: Identification of cellular proteins that interact with Newcastle Disease Virus and human Respiratory Syncytial Virus by a two-dimensional virus overlay protein binding assay (VOPBA).
abstract: Although it is well documented that the initial attachment receptors for Newcastle Disease Virus (NDV) and Respiratory Syncytial Virus (RSV) are sialic acid-containing molecules and glycosaminoglycans respectively, the exact nature of the receptors for both viruses remains to be deciphered. Moreover, additional molecules at the host cell surface might be involved in the entry mechanism. With the aim of identifying the cellular proteins that interact with NDV and RSV at the cell surface, we performed a virus overlay protein binding assay (VOPBA). Cell membrane lysates were separated by two dimensional (2D) gel electrophoresis and electrotransferred to PVDF membranes, after which they were probed with high viral concentrations. NDV interacted with a Protein Disulfide Isomerase from chicken fibroblasts. In the case of RSV, we detected 15 reactive spots, which were identified as six different proteins, of which nucleolin was outstanding. We discuss the possible role of PDI and nucleolin in NDV and RSV entry, respectively.
authors.affiliation: Departamento de Bioquímica y Biología Molecular, Universidad de Salamanca, Edificio Departamental Lab. 106, Plaza Doctores de la Reina s/n, 37007 Salamanca, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Holguera
authors.affiliation: Departamento de Bioquímica y Biología Molecular, Universidad de Salamanca, Edificio Departamental Lab. 106, Plaza Doctores de la Reina s/n, 37007 Salamanca, Spain. Electronic address: evillar@usal.es.
authors.forename: Enrique
authors.initials: E
authors.lastname: Villar
authors.affiliation: Departamento de Bioquímica y Biología Molecular, Universidad de Salamanca, Edificio Departamental Lab. 106, Plaza Doctores de la Reina s/n, 37007 Salamanca, Spain. Electronic address: imunbar@usal.es.
authors.forename: Isabel
authors.initials: I
authors.lastname: Muñoz-Barroso
mesh_terms: D000818:Animals; D002460:Cell Line; D002645:Chickens; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D009521:Newcastle Disease / Q000378:metabolism* / Q000821:virology; D009522:Newcastle disease virus / Q000235:genetics / Q000378:metabolism*; D011201:Poultry Diseases / Q000378:metabolism* / Q000821:virology; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018357:Respiratory Syncytial Virus Infections / Q000378:metabolism* / Q000821:virology; D018113:Respiratory Syncytial Virus, Human / Q000235:genetics / Q000378:metabolism*
chemical_list: D011506:Proteins
keywords: NDV; Nucleolin; PDI; RSV; VOPBA