title: Purification and characterization of BmooAi: a new toxin from Bothrops moojeni snake venom that inhibits platelet aggregation.
abstract: In this paper, we describe the purification/characterization of BmooAi, a new toxin from Bothrops moojeni that inhibits platelet aggregation. The purification of BmooAi was carried out through three chromatographic steps (ion-exchange on a DEAE-Sephacel column, molecular exclusion on a Sephadex G-75 column, and reverse-phase HPLC chromatography on a C2/C18 column). BmooAi was homogeneous by SDS-PAGE and shown to be a single-chain protein of 15,000 Da. BmooAi was analysed by MALDI-TOF Spectrometry and revealed two major components with molecular masses 7824.4 and 7409.2 as well as a trace of protein with a molecular mass of 15,237.4 Da. Sequencing of BmooAi by Edman degradation showed two amino acid sequences: IRDFDPLTNAPENTA and ETEEGAEEGTQ, which revealed no homology to any known toxin from snake venom. BmooAi showed a rather specific inhibitory effect on platelet aggregation induced by collagen, adenosine diphosphate, or epinephrine in human platelet-rich plasma in a dose-dependent manner, whereas it had little or no effect on platelet aggregation induced by ristocetin. The effect on platelet aggregation induced by BmooAi remained active even when heated to 100°C. BmooAi could be of medical interest as a new tool for the development of novel therapeutic agents for the prevention and treatment of thrombotic disorders.
authors: Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, 38400-902 Uberlândia, MG, Brazil ; Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica (N-Biofar), 31270-901 Belo Horizonte, MG, Brazil.
authors: Mayara Ribeiro
authors: MR
authors: de Queiroz
authors: Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, 38400-902 Uberlândia, MG, Brazil ; Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica (N-Biofar), 31270-901 Belo Horizonte, MG, Brazil.
authors: Carla Cristine N
authors: CC
authors: Mamede
authors: Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, 38400-902 Uberlândia, MG, Brazil ; Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica (N-Biofar), 31270-901 Belo Horizonte, MG, Brazil.
authors: Nadia Cristina G
authors: NC
authors: de Morais
authors: Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, 38400-902 Uberlândia, MG, Brazil ; Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica (N-Biofar), 31270-901 Belo Horizonte, MG, Brazil.
authors: Kelly Cortes
authors: KC
authors: Fonseca
authors: Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, 38400-902 Uberlândia, MG, Brazil ; Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica (N-Biofar), 31270-901 Belo Horizonte, MG, Brazil.
authors: Bruna Barbosa
authors: BB
authors: de Sousa
authors: Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, 38400-902 Uberlândia, MG, Brazil.
authors: Thaís M
authors: TM
authors: Migliorini
authors: Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, 38400-902 Uberlândia, MG, Brazil.
authors: Déborah Fernanda C
authors: DF
authors: Pereira
authors: Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica (N-Biofar), 31270-901 Belo Horizonte, MG, Brazil ; Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, 38400-902 Uberlândia, MG, Brazil.
authors: Leonilda
authors: L
authors: Stanziola
authors: Centro de Estudos de Biomoléculas Aplicadas à Saúde (CEBio), Fundação Oswaldo Cruz Rondônia (Fiocruz Rondônia) e Núcleo de Saúde, Universidade Federal de Rondônia (UNIR), 76812-245 Porto Velho, RO, Brazil.
authors: Leonardo A
authors: LA
authors: Calderon
authors: Centro de Estudos de Biomoléculas Aplicadas à Saúde (CEBio), Fundação Oswaldo Cruz Rondônia (Fiocruz Rondônia) e Núcleo de Saúde, Universidade Federal de Rondônia (UNIR), 76812-245 Porto Velho, RO, Brazil.
authors: Rodrigo
authors: R
authors: Simões-Silva
authors: Centro de Estudos de Biomoléculas Aplicadas à Saúde (CEBio), Fundação Oswaldo Cruz Rondônia (Fiocruz Rondônia) e Núcleo de Saúde, Universidade Federal de Rondônia (UNIR), 76812-245 Porto Velho, RO, Brazil.
authors: Andreimar Martins
authors: AM
authors: Soares
authors: Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica (N-Biofar), 31270-901 Belo Horizonte, MG, Brazil ; Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, 38400-902 Uberlândia, MG, Brazil.
authors: Fábio
authors: F
authors: de Oliveira
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017837:Bothrops / Q000378:metabolism*; D002417:Cattle; D002852:Chromatography, Ion Exchange; D003094:Collagen / Q000494:pharmacology; D003435:Crotalid Venoms / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D004837:Epinephrine / Q000494:pharmacology; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010974:Platelet Aggregation / Q000187:drug effects*; D010975:Platelet Aggregation Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D003435:Crotalid Venoms; D010975:Platelet Aggregation Inhibitors; D003094:Collagen; D004837:Epinephrine