title: Molecular and functional characterization of metalloserrulases, new metalloproteases from the Tityus serrulatus venom gland.
abstract: Tityus serrulatus is a Brazilian scorpion species with great medical significance. While the effects of neurotoxins have been extensively studied, little is known about the proteases expressed in the venom gland of this arthropod. In this study, clones from a T. serrulatus (Ts) venom gland cDNA library were selected according to homology to proteases. The sequences were aligned in the database and classified by homology. Similarity and identity analyses of the sequences were carried out, and a phylogenetic tree was constructed with the sequences of other proteases. These cDNA sequences correspond to ten different metalloproteases, named metalloserrulases (TsMS). TsMS 1-9 belong to the metzincin family, which has three domains: signal peptide, propeptide, and metalloprotease domain; while TsMS 10 belongs to the gluzincin family. The proteolytic activity of the venom was inferred from the cleavage of fibrinogen, and the residues recognized by the proteases were determined by cleavage of a tripeptide library using a fluorescence resonance energy transfer assay. The Ts venom showed proteolytic activity on fibrinogen and preferential cleavage close to the basic residues K and R. Its activity could be inhibited by EDTA, indicating that the venom from this scorpion predominantly consists of metalloproteases.
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Presidente Antônio Carlos, 6627 - Pampulha, Belo Horizonte, MG, 31270-901, Brazil. Electronic address: anderson.o.carmo@gmail.com.
authors.forename: A O
authors.initials: AO
authors.lastname: Carmo
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Presidente Antônio Carlos, 6627 - Pampulha, Belo Horizonte, MG, 31270-901, Brazil. Electronic address: barbarabio2@gmail.com.
authors.forename: B B R
authors.initials: BB
authors.lastname: Oliveira-Mendes
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Presidente Antônio Carlos, 6627 - Pampulha, Belo Horizonte, MG, 31270-901, Brazil. Electronic address: carolinacampolina@yahoo.com.br.
authors.forename: C C R
authors.initials: CC
authors.lastname: Horta
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Presidente Antônio Carlos, 6627 - Pampulha, Belo Horizonte, MG, 31270-901, Brazil. Electronic address: barbara_fmag@hotmail.com.
authors.forename: B F
authors.initials: BF
authors.lastname: Magalhães
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Presidente Antônio Carlos, 6627 - Pampulha, Belo Horizonte, MG, 31270-901, Brazil. Electronic address: arthurestanislau@gmail.com.
authors.forename: A E
authors.initials: AE
authors.lastname: Dantas
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Presidente Antônio Carlos, 6627 - Pampulha, Belo Horizonte, MG, 31270-901, Brazil. Electronic address: lucasdemeirachaves@hotmail.com.
authors.forename: L M
authors.initials: LM
authors.lastname: Chaves
authors.affiliation: Departamento de Bioquímica - Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Presidente Antônio Carlos, 6627 - Pampulha, Belo Horizonte, MG, 31270-901, Brazil. Electronic address: olortegi@icb.ufmg.br.
authors.forename: C
authors.initials: C
authors.lastname: Chávez-Olórtegui
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Presidente Antônio Carlos, 6627 - Pampulha, Belo Horizonte, MG, 31270-901, Brazil. Electronic address: ekalapo@icb.ufmg.br.
authors.forename: E
authors.initials: E
authors.lastname: Kalapothakis
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D018076:DNA, Complementary / Q000235:genetics; D005340:Fibrinogen / Q000378:metabolism; D045726:Metalloproteases / Q000737:chemistry / Q000235:genetics* / Q000633:toxicity*; D008969:Molecular Sequence Data; D010802:Phylogeny; D012604:Scorpion Venoms / Q000201:enzymology*; D012605:Scorpions; D017386:Sequence Homology, Amino Acid
chemical_list: D018076:DNA, Complementary; D012604:Scorpion Venoms; D005340:Fibrinogen; D045726:Metalloproteases
keywords: Envenomation; Metalloprotease; Metalloserrulase; Scorpion; Tityus serrulatus; Venom