title: Molecular characterization of metalloproteases from Bothrops alternatus snake venom.
abstract: We have previously demonstrated that alternagin-C (ALT-C), a disintegrin-like, Cys-rich protein isolated from Bothrops alternatus snake venom, induces human vascular endothelial cell (HUVEC) proliferation and angiogenesis in in vitro and in vivo assays. Therefore this protein could be interesting as a new approach for tissue regeneration studies. However, its primary sequence was not completely determined since the protein isolated from crude venom is usually a mixture of isoforms. Here we describe the transcriptome analysis of B. alternatus from the venom glands of a single male specimen. About 800 good-quality contigs were screened for snake venom metalloproteases/disintegrins, resulting in the following expression profile for these enzymes: 4% for P-I, 7% for P-II and 89% for P-III SVMPs. The PII-SVMP sequence code for RGD-disintegrins and all the expressed PIII-sequences have the ECD adhesive motif. A cDNA sequence coding for an ALT-C homolog was completely sequenced and characterized. Comparative sequence and structural analyses suggested new features that distinguish SVMP classes such as two prolyl endopetidase cleavage sites. All these data add new information on the expression pattern of metalloproteases of B. alternatus venom and may have practical applications for the production of recombinant disintegrins for cell adhesion studies.
authors.affiliation: Departamento de Genética e Evolução, Universidade Federal de São Carlos, São Carlos, SP, Brazil; Universidade de Brasília, Campus Planaltina - UnB/FUP Brazil, Brazil.
authors.forename: Fernando Fonseca Pereira
authors.initials: FF
authors.lastname: de Paula
authors.affiliation: Departamento de Ciências Fisiológicas, Universidade Federal de São Carlos, São Carlos, SP, Brazil.
authors.forename: Juliana Uema
authors.initials: JU
authors.lastname: Ribeiro
authors.affiliation: Departamento de Ciências Fisiológicas, Universidade Federal de São Carlos, São Carlos, SP, Brazil.
authors.forename: Livia Mara
authors.initials: LM
authors.lastname: Santos
authors.affiliation: Departamento de Química, Universidade Federal de São Carlos, São Carlos, SP, Brazil.
authors.forename: Dulce Helena Ferreira
authors.initials: DH
authors.lastname: de Souza
authors.affiliation: Departamento de Genética e Evolução, Universidade Federal de São Carlos, São Carlos, SP, Brazil; EMBRAPA Genetic Resources and Biotechnology, Brazil.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Leonardecz
authors.affiliation: Departamento de Genética e Evolução, Universidade Federal de São Carlos, São Carlos, SP, Brazil.
authors.forename: Flávio
authors.initials: F
authors.lastname: Henrique-Silva
authors.affiliation: Departamento de Ciências Fisiológicas, Universidade Federal de São Carlos, São Carlos, SP, Brazil. Electronic address: hsaraujo@ufscar.br.
authors.forename: Heloisa Sobreiro
authors.initials: HS
authors.lastname: Selistre-de-Araújo
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017837:Bothrops / Q000235:genetics / Q000378:metabolism; D003435:Crotalid Venoms / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D019483:Disintegrins / Q000737:chemistry / Q000235:genetics; D008297:Male; D045726:Metalloproteases / Q000737:chemistry* / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D059467:Transcriptome
chemical_list: D003435:Crotalid Venoms; D019483:Disintegrins; C481346:alternagin-C; D045726:Metalloproteases
keywords: Alternagin-C; Bothrops alternatus; Disintegrin; Transcriptome; Venom gland