title: Identification of protective B-cell epitopes of Atroxlysin-I: A metalloproteinase from Bothrops atrox snake venom.
abstract: Atroxlysin-I (Atr-I) is a hemorrhagic snake venom metalloproteinase (SVMP) from Bothrops atrox venom, the snake responsible for the majority of bites in the north region of South America. SVMPs like Atr-I produce toxic effects in victims including hemorrhage, inflammation, necrosis and blood coagulation deficiency. Mapping of B-cell epitopes in SVMPs might result in the identification of non-toxic molecules capable of inducing neutralizing antibodies and improving the anti-venom therapy. Here, using the SPOT-synthesis technique we identified two epitopes located in the N-ter region of Atr-I (AtrEp1-(22)YNGNSDKIRRRIHQM(36); and AtrEp2-(55)GVEIWSNKDLINVQ(68)). Based on the sequence of AtrEp1 and AtrEp2 a third peptide named Atr-I biepitope (AtrBiEp) was designed and synthesized ((23)NGNSDKIRRRIH(34)GG(55)GVEIWSNKDLINVQ(68)). AtrBiEp was used to immunize BALB/c mice. Anti-AtrBiEp serum cross-reacted against Atr-I in western blot and was able to fully neutralize the hemorrhagic activity of Atr-I. Our results provide a rational basis for the identification of neutralizing epitopes on Atr-I snake venom toxin and show that the use of synthetic peptides could improve the generation of immuno-therapeutics.
authors.affiliation: Departamento de Bioquímica e Imunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil; Sys2Diag, FRE 3690, CNRS Alcediag, Languedoc-Roussillon, Montpellier, France.
authors.forename: F S
authors.initials: FS
authors.lastname: Schneider
authors.affiliation: Departamento de Bioquímica e Imunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
authors.forename: S
authors.initials: S
authors.lastname: de Almeida Lima
authors.affiliation: Departamento de Bioquímica e Imunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
authors.forename: G
authors.initials: G
authors.lastname: Reis de Ávila
authors.affiliation: Departamento de Bioquímica e Imunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
authors.forename: K L
authors.initials: KL
authors.lastname: Castro
authors.affiliation: Departamento de Bioquímica e Imunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
authors.forename: C
authors.initials: C
authors.lastname: Guerra-Duarte
authors.affiliation: Diretoria de Pesquisa e Desenvolvimento, Fundação Ezequiel Dias, Belo Horizonte, MG, Brazil.
authors.forename: E F
authors.initials: EF
authors.lastname: Sanchez
authors.affiliation: Sys2Diag, FRE 3690, CNRS Alcediag, Languedoc-Roussillon, Montpellier, France.
authors.forename: C
authors.initials: C
authors.lastname: Nguyen
authors.affiliation: Sys2Diag, FRE 3690, CNRS Alcediag, Languedoc-Roussillon, Montpellier, France.
authors.forename: C
authors.initials: C
authors.lastname: Granier
authors.affiliation: Sys2Diag, FRE 3690, CNRS Alcediag, Languedoc-Roussillon, Montpellier, France.
authors.forename: F
authors.initials: F
authors.lastname: Molina
authors.affiliation: Departamento de Bioquímica e Imunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil. Electronic address: olortegi@icb.ufmg.br.
authors.forename: C
authors.initials: C
authors.lastname: Chávez-Olortegui
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D057134:Antibodies, Neutralizing / Q000276:immunology; D000997:Antivenins / Q000276:immunology*; D017837:Bothrops; D003429:Cross Reactions; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D008666:Metalloendopeptidases / Q000276:immunology*; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D010455:Peptides / Q000276:immunology; D017434:Protein Structure, Tertiary; D012910:Snake Venoms / Q000276:immunology*
chemical_list: D057134:Antibodies, Neutralizing; D000997:Antivenins; D018985:Epitopes, B-Lymphocyte; D010455:Peptides; D012910:Snake Venoms; D008666:Metalloendopeptidases; C069317:hemorrhagic metalloproteinase
keywords: Hemorrhage; Linear B-cell epitope mapping; Peptide synthesis; Snake venom metalloproteinases