title: Alkylation of histidine residues of Bothrops jararacussu venom proteins and isolated phospholipases A2: a biotechnological tool to improve the production of antibodies.
abstract: Crude venom of Bothrops jararacussu and isolated phospholipases A2 (PLA2) of this toxin (BthTX-I and BthTX-II) were chemically modified (alkylation) by p-bromophenacyl bromide (BPB) in order to study antibody production capacity in function of the structure-function relationship of these substances (crude venom and PLA2 native and alkylated). BthTX-II showed enzymatic activity, while BthTX-I did not. Alkylation reduced BthTX-II activity by 50% while this process abolished the catalytic and myotoxic activities of BthTX-I, while reducing its edema-inducing activity by about 50%. Antibody production against the native and alkylated forms of BthTX-I and -II and the cross-reactivity of antibodies to native and alkylated toxins did not show any apparent differences and these observations were reinforced by surface plasmon resonance (SPR) data. Histopathological analysis of mouse gastrocnemius muscle sections after injection of PBS, BthTX-I, BthTX-II, or both myotoxins previously incubated with neutralizing antibody showed inhibition of the toxin-induced myotoxicity. These results reveal that the chemical modification of the phospholipases A2 (PLA2) diminished their toxicity but did not alter their antigenicity. This observation indicates that the modified PLA2 may provide a biotechnological tool to attenuate the toxicity of the crude venom, by improving the production of antibodies and decreasing the local toxic effects of this poisonous substance in animals used to produce antivenom.
authors.affiliation: Center of Applied Biomolecules Studies in Health (CEBio), Oswaldo Cruz Foundation (Fiocruz Rondônia), Porto Velho, RO, Brazil ; Department of Medicine, Federal University of Rondônia (UNIR), Porto Velho, RO, Brazil ; Brazilian Institute of Environment and Renewable Natural Resources (IBAMA), Porto Velho, RO, Brazil.
authors.forename: C L S
authors.initials: CL
authors.lastname: Guimarães
authors.affiliation: Department of Biochemistry and Immunology, Faculty of Medicine of Ribeirão Preto, University of São Paulo (USP), Ribeirão Preto, SP, Brazil.
authors.forename: S H
authors.initials: SH
authors.lastname: Andrião-Escarso
authors.affiliation: Center of Applied Biomolecules Studies in Health (CEBio), Oswaldo Cruz Foundation (Fiocruz Rondônia), Porto Velho, RO, Brazil ; Department of Medicine, Federal University of Rondônia (UNIR), Porto Velho, RO, Brazil.
authors.forename: L S
authors.initials: LS
authors.lastname: Moreira-Dill
authors.affiliation: Department of Chemistry, Biotechnology and Bioprocess Engineering, Federal University of São João Del Rei (UFSJ), Ouro Branco, MG, Brazil.
authors.forename: B M A
authors.initials: BM
authors.lastname: Carvalho
authors.affiliation: Department of Molecular Biology, Center for Science and Nature, Federal University of Paraíba (UFPB), João Pessoa, Brazil.
authors.forename: D P
authors.initials: DP
authors.lastname: Marchi-Salvador
authors.affiliation: Department of Clinical Analysis, Faculty of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo (USP), Ribeirão Preto, SP, Brazil.
authors.forename: N A
authors.initials: NA
authors.lastname: Santos-Filho
authors.affiliation: Department of Physics and Biophysics, State University Paulista (UNESP), Botucatu, SP, Brazil.
authors.forename: C A H
authors.initials: CA
authors.lastname: Fernandes
authors.affiliation: Department of Physics and Biophysics, State University Paulista (UNESP), Botucatu, SP, Brazil.
authors.forename: M R M
authors.initials: MR
authors.lastname: Fontes
authors.affiliation: Department of Biochemistry and Immunology, Faculty of Medicine of Ribeirão Preto, University of São Paulo (USP), Ribeirão Preto, SP, Brazil.
authors.forename: J R
authors.initials: JR
authors.lastname: Giglio
authors.affiliation: Center for the Study of Venoms and Venomous Animals (CEVAP), State University Paulista (UNESP), Botucatu, SP, Brazil.
authors.forename: B
authors.initials: B
authors.lastname: Barraviera
authors.affiliation: Center of Applied Biomolecules Studies in Health (CEBio), Oswaldo Cruz Foundation (Fiocruz Rondônia), Porto Velho, RO, Brazil ; Department of Medicine, Federal University of Rondônia (UNIR), Porto Velho, RO, Brazil.
authors.forename: J P
authors.initials: JP
authors.lastname: Zuliani
authors.affiliation: Center of Applied Biomolecules Studies in Health (CEBio), Oswaldo Cruz Foundation (Fiocruz Rondônia), Porto Velho, RO, Brazil ; Department of Medicine, Federal University of Rondônia (UNIR), Porto Velho, RO, Brazil.
authors.forename: C F C
authors.initials: CF
authors.lastname: Fernandes
authors.affiliation: Center of Applied Biomolecules Studies in Health (CEBio), Oswaldo Cruz Foundation (Fiocruz Rondônia), Porto Velho, RO, Brazil ; Department of Medicine, Federal University of Rondônia (UNIR), Porto Velho, RO, Brazil.
authors.forename: L A
authors.initials: LA
authors.lastname: Calderón
authors.affiliation: Center of Applied Biomolecules Studies in Health (CEBio), Oswaldo Cruz Foundation (Fiocruz Rondônia), Porto Velho, RO, Brazil ; Department of Medicine, Federal University of Rondônia (UNIR), Porto Velho, RO, Brazil.
authors.forename: R G
authors.initials: RG
authors.lastname: Stábeli
authors.affiliation: Institute for Research in Biomedicine (IRB Barcelona) and CIBER-BBN, Barcelona Science Park, Barcelona, Spain ; Department of Organic Chemistry, University of Barcelona, Barcelona, Spain ; School of Chemistry and Physics, University of KwaZulu Natal, Durban 4001, South Africa.
authors.forename: F
authors.initials: F
authors.lastname: Albericio
authors.affiliation: Department of Chemistry, Biotechnology and Bioprocess Engineering, Federal University of São João Del Rei (UFSJ), Ouro Branco, MG, Brazil.
authors.forename: S L
authors.initials: SL
authors.lastname: da Silva
authors.affiliation: Center of Applied Biomolecules Studies in Health (CEBio), Oswaldo Cruz Foundation (Fiocruz Rondônia), Porto Velho, RO, Brazil ; Department of Medicine, Federal University of Rondônia (UNIR), Porto Velho, RO, Brazil.
authors.forename: A M
authors.initials: AM
authors.lastname: Soares
mesh_terms: D000478:Alkylation / Q000276:immunology*; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000997:Antivenins / Q000276:immunology / Q000378:metabolism; D017837:Bothrops / Q000276:immunology / Q000378:metabolism*; D003429:Cross Reactions / Q000276:immunology; D003435:Crotalid Venoms / Q000276:immunology / Q000378:metabolism*; D006639:Histidine / Q000276:immunology / Q000378:metabolism*; D008297:Male; D051379:Mice; D018482:Muscle, Skeletal / Q000276:immunology / Q000378:metabolism; D054467:Phospholipases A2 / Q000276:immunology / Q000378:metabolism*
chemical_list: D000906:Antibodies; D000997:Antivenins; D003435:Crotalid Venoms; D006639:Histidine; D054467:Phospholipases A2