Venomic, Transcriptomic, and Bioactivity Analyses of Pamphobeteus verdolaga Venom Reveal Complex Disulfide-Rich Peptides That Modulate Calcium Channels

Pamphobeteus verdolaga is a recently described Theraphosidae spider from the Andean region of Colombia. Previous reports partially characterized its venom profile. In this study, we conducted a detailed analysis that includes reversed-phase high-performance liquid chromatography (rp-HPLC), calcium i...

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Autores:
Estrada Gomez, Sebastian
Caldas Cardoso, Fernanda
Vargas Muñoz, Leidy Johana
Quintana Castillo, Juan Carlos
Arenas Gómez, Claudia Marcela
Pineda, Sandy Steffany
Saldarriaga Cordoba, Monica Maria
Tipo de recurso:
Article of journal
Fecha de publicación:
2019
Institución:
Universidad Cooperativa de Colombia
Repositorio:
Repositorio UCC
Idioma:
OAI Identifier:
oai:repository.ucc.edu.co:20.500.12494/41485
Acceso en línea:
https://doi.org/10.1016/j.rcp.2014.02.004
https://hdl.handle.net/20.500.12494/41485
Palabra clave:
Pamphobeteus
disulfide-rich peptide (DRP)
inhibitory cysteine knot (ICK)
ion channels
peptides
theraphosidae
transcriptome
venomics
Rights
closedAccess
License
http://purl.org/coar/access_right/c_14cb
Description
Summary:Pamphobeteus verdolaga is a recently described Theraphosidae spider from the Andean region of Colombia. Previous reports partially characterized its venom profile. In this study, we conducted a detailed analysis that includes reversed-phase high-performance liquid chromatography (rp-HPLC), calcium influx assays, tandem mass spectrometry analysis (tMS/MS), and venom-gland transcriptome. rp-HPLC fractions of P. verdolaga venom showed activity on Ca V 2.2, Ca V 3.2, and Na V 1.7 ion channels. Active fractions contained several peptides with molecular masses ranging from 3399.4 to 3839.6 Da. The tMS/MS analysis of active fraction displaying the strongest activity to inhibit calcium channels showed sequence fragments similar to one of the translated transcripts detected in the venom-gland transcriptome. The putative peptide of this translated transcript corresponded to a toxin, here named ?-theraphositoxin-Pv3a, a potential ion channel modulator toxin that is, in addition, very similar to other theraphositoxins affecting calcium channels (i.e., ?-theraphotoxin-Asp1a). Additionally, using this holistic approach, we found that P. verdolaga venom is an important source of disulfide-rich proteins expressing at least eight superfamilies.