title: Proteomic profiling of the infective trophozoite stage of Acanthamoeba polyphaga.
abstract: Acanthamoeba polyphaga is a free-living protozoan pathogen, whose infective trophozoite form is capable of causing a blinding keratitis and fatal granulomatous encephalitis in humans. The damage caused by A. polyphaga trophozoites in human corneal or brain infections is the result of several different pathogenic mechanisms that have not yet been elucidated at the molecular level. We performed a comprehensive analysis of the proteins expressed by A. polyphaga trophozoites, based on complementary 2-DE MS/MS and gel-free LC-MS/MS approaches. Overall, 202 non-redundant proteins were identified. An A. polyphaga proteomic map in the pH range 3-10 was produced, with protein identification for 184 of 370 resolved spots, corresponding to 142 proteins. Additionally, 94 proteins were identified by gel-free LC-MS/MS. Functional classification revealed several proteins with potential importance for pathogen survival and infection of mammalian hosts, including surface proteins and proteins related to defense mechanisms. Our study provided the first comprehensive proteomic survey of the trophozoite infective stage of an Acanthamoeba species, and established foundations for prospective, comparative and functional studies of proteins involved in mechanisms of survival, development, and pathogenicity in A. polyphaga and other pathogenic amoebae.
authors.affiliation: Laboratório de Parasitologia, Instituto de Ciências Básicas da Saúde, Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal do Rio Grande do Sul, CEP: 90050170 Porto Alegre, RS, Brazil. Electronic address: k.caumo@ufsc.br.
authors.forename: Karin Silva
authors.initials: KS
authors.lastname: Caumo
authors.affiliation: Laboratório de Genômica Estrutural e Funcional, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, CEP: 91501-970 Porto Alegre, RS, Brazil. Electronic address: karina@cbiot.ufrgs.br.
authors.forename: Karina Mariante
authors.initials: KM
authors.lastname: Monteiro
authors.affiliation: Laboratório de Genômica Estrutural e Funcional, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, CEP: 91501-970 Porto Alegre, RS, Brazil. Electronic address: thiely.ott@gmail.com.
authors.forename: Thiely Rodrigues
authors.initials: TR
authors.lastname: Ott
authors.affiliation: Laboratório de Parasitologia, Instituto de Ciências Básicas da Saúde, Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal do Rio Grande do Sul, CEP: 90050170 Porto Alegre, RS, Brazil. Electronic address: vinimaschio@yahoo.com.br.
authors.forename: Vinicius José
authors.initials: VJ
authors.lastname: Maschio
authors.affiliation: Laboratório de Doenças Infecciosas e Parasitárias, Universidade do Oeste de Santa Catarina, CEP: 89600-000 Joaçaba, SC, Brazil. Electronic address: glauberwagner@gmail.com.
authors.forename: Glauber
authors.initials: G
authors.lastname: Wagner
authors.affiliation: Laboratório de Genômica Estrutural e Funcional, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, CEP: 91501-970 Porto Alegre, RS, Brazil. Electronic address: henrique@cbiot.ufrgs.br.
authors.forename: Henrique Bunselmeyer
authors.initials: HB
authors.lastname: Ferreira
authors.affiliation: Laboratório de Parasitologia, Instituto de Ciências Básicas da Saúde, Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal do Rio Grande do Sul, CEP: 90050170 Porto Alegre, RS, Brazil. Electronic address: marilise.rott@ufrgs.br.
authors.forename: Marilise Brittes
authors.initials: MB
authors.lastname: Rott
mesh_terms: D000048:Acanthamoeba / Q000378:metabolism*; D015823:Acanthamoeba Keratitis / Q000469:parasitology* / Q000517:prevention & control; D000818:Animals; D006801:Humans; D040901:Proteomics; D053719:Tandem Mass Spectrometry; D053058:Trophozoites / Q000378:metabolism
keywords: 2-DE; Acanthamoeba; Global protein analysis; LC–MS/MS; Pathogen–host interaction; Trophozoite