title: Peptidomics of Acanthoscurria gomesiana spider venom reveals new toxins with potential antimicrobial activity.
abstract: Acanthoscurria gomesiana is a Brazilian spider from the Theraphosidae family inhabiting regions of Southeastern Brazil. Potent antimicrobial peptides as gomesin and acanthoscurrin have been discovered from the spider hemolymph in previous works. Spider venoms are also recognized as sources of biologically active peptides, however the venom peptidome of A. gomesiana remained unexplored to date. In this work, a MS-based workflow was applied to the investigation of the spider venom peptidome. Data-independent and data-dependent LC-MS/MS acquisitions of intact peptides and of peptides submitted to multiple enzyme digestions, followed by automated chromatographic alignment, de novo analysis, database and homology searches with manual validations showed that the venom is composed by <165 features, with masses ranging from 0.4-15.8kDa. From digestions, 135 peptides were identified from 17 proteins, including three new mature peptides: U1-TRTX-Agm1a, U1-TRTX-Agm2a and U1-TRTX-Agm3a, containing 3, 4 and 3 disulfide bonds, respectively. The toxins U1-TRTX-Agm1a differed by only one amino acid from U1-TRTX-Ap1a from A. paulensis and U1-TRTX-Agm2a was derived from the genicutoxin-D1 precursor from A. geniculata. These toxins have potential applications as antimicrobial agents, as the peptide fraction of A. gomesiana showed activity against Escherichia coli, Enterobacter cloacae and Candida albicans strains. MS data are available via ProteomeXchange Consortium with identifier PXD003884.
  
  
  
  Biological fluids of the Acanthoscurria gomesiana spider are sources of active molecules, as is the case of antimicrobial peptides and acylpolyamines found in the hemolymphs. The venom is also a potential source of toxins with pharmacological and biotechnological applications. However, to our knowledge no A. gomesiana venom toxin structure has been determined to date. Using a combination of high resolution mass spectrometry, transcriptomics and bioinformatics, we employed a workflow to fully sequence, determine the number of disulfide bonds of mature peptides and we found new potential antimicrobial peptides. This workflow is suitable for complete peptide toxin sequencing when handling limited amount of venom samples and can accelerate the discovery of peptides with potential biotechnological applications.
authors.affiliation: Departamento de Bioquímica, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP, Brazil.
authors.forename: Thiago F
authors.initials: TF
authors.lastname: Abreu
authors.affiliation: Departamento de Bioquímica, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP, Brazil.
authors.forename: Bianca N
authors.initials: BN
authors.lastname: Sumitomo
authors.affiliation: Laboratório Especial de Toxinologia Aplicada, Center of Toxins, Immune-Response and Cell Signaling, Instituto Butantan, São Paulo, SP, Brazil.
authors.forename: Milton Y
authors.initials: MY
authors.lastname: Nishiyama
authors.affiliation: Laboratório Especial de Toxinologia Aplicada, Center of Toxins, Immune-Response and Cell Signaling, Instituto Butantan, São Paulo, SP, Brazil.
authors.forename: Ursula C
authors.initials: UC
authors.lastname: Oliveira
authors.affiliation: Mass Spectrometry Applications Research & Development Laboratory, Waters Corporation, Sāo Paulo, SP, Brazil.
authors.forename: Gustavo H M F
authors.initials: GH
authors.lastname: Souza
authors.affiliation: Laboratório Especial de Toxinologia Aplicada, Center of Toxins, Immune-Response and Cell Signaling, Instituto Butantan, São Paulo, SP, Brazil.
authors.forename: Eduardo S
authors.initials: ES
authors.lastname: Kitano
authors.affiliation: Departamento de Ciência e Tecnologia, Universidade Federal de São Paulo, ICT-UNIFESP, São José dos Campos, SP, Brazil.
authors.forename: André
authors.initials: A
authors.lastname: Zelanis
authors.affiliation: Laboratório Especial de Toxinologia Aplicada, Center of Toxins, Immune-Response and Cell Signaling, Instituto Butantan, São Paulo, SP, Brazil.
authors.forename: Solange M T
authors.initials: SM
authors.lastname: Serrano
authors.affiliation: Laboratório Especial de Toxinologia Aplicada, Center of Toxins, Immune-Response and Cell Signaling, Instituto Butantan, São Paulo, SP, Brazil.
authors.forename: Inácio
authors.initials: I
authors.lastname: Junqueira-de-Azevedo
authors.affiliation: Laboratório Especial de Toxinologia Aplicada, Center of Toxins, Immune-Response and Cell Signaling, Instituto Butantan, São Paulo, SP, Brazil.
authors.forename: Pedro I
authors.initials: PI
authors.lastname: Silva
authors.affiliation: Departamento de Bioquímica, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP, Brazil. Electronic address: aktashima@unifesp.br.
authors.forename: Alexandre K
authors.initials: AK
authors.lastname: Tashima
mesh_terms: D000818:Animals; D000890:Anti-Infective Agents / Q000302:isolation & purification* / Q000494:pharmacology; D001938:Brazil; D002853:Chromatography, Liquid; D004220:Disulfides / Q000032:analysis / Q000494:pharmacology; D010446:Peptide Fragments / Q000032:analysis / Q000494:pharmacology; D010455:Peptides / Q000032:analysis* / Q000494:pharmacology; D013111:Spider Venoms / Q000737:chemistry*; D013112:Spiders / Q000472:pathogenicity*; D053719:Tandem Mass Spectrometry; D057188:Workflow
chemical_list: D000890:Anti-Infective Agents; D004220:Disulfides; D010446:Peptide Fragments; D010455:Peptides; D013111:Spider Venoms
keywords: Acanthoscurria gomesiana; Antimicrobial peptide; Peptidomics; Theraphotoxin