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Open AccessArticle

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

1
Programa de Ofidismo/Escorpionismo—Serpentario, Universidad de Antioquia UdeA, Carrera 53 No 61–30, Medellín, Antioquia, CO 050010, Colombia
2
Facultad de Ciencias Farmacéuticas y Alimentarias, Universidad de Antioquia UdeA, Calle 70 No 52–21, Medellín, Antioquia, CO 050010, Colombia
3
Institute for Molecular Bioscience, The University of Queensland, 306 Carmody Road, St Lucia, QLD 4072, Australia
4
Facultad de Medicina, Universidad Cooperativa de Colombia, Calle 50 A No 41–20 Medellín, Antioquia, CO 050012, Colombia
5
Grupo de Génetica, Regeneración y Cáncer, Universidad de Antioquia UdeA, Carrera 53 No 61–30, Medellín, Antioquia, CO 050010, Colombia
6
Brain and Mind Centre, University of Sydney, Camperdown, NSW 2052, Australia
7
Garvan Institute of Medical Research, Darlinghurst, Sydney, NSW 2010, Australia
8
St. Vincent’s Clinical School, University of New South Wales, Sydney, NSW 2010, Australia
9
Centro de Investigación en Recursos Naturales y Sustentabilidad, Universidad Bernardo O’Higgins, Avenida Viel 1497, Santiago 7750000, Chile
*
Author to whom correspondence should be addressed.
Current address: Marine Biological Laboratory, Eugene Bell Center for Regenerative Biology and Tissue Engineering, Woods Hole, MA 02543, USA.
Toxins 2019, 11(9), 496; https://doi.org/10.3390/toxins11090496
Received: 23 June 2019 / Revised: 12 August 2019 / Accepted: 14 August 2019 / Published: 27 August 2019
(This article belongs to the Special Issue Venoms and Ion 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 influx assays, tandem mass spectrometry analysis (tMS/MS), and venom-gland transcriptome. rp-HPLC fractions of P. verdolaga venom showed activity on CaV2.2, CaV3.2, and NaV1.7 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 ω-theraphositoxin-Pv3a, a potential ion channel modulator toxin that is, in addition, very similar to other theraphositoxins affecting calcium channels (i.e., ω-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. View Full-Text
Keywords: theraphosidae; Pamphobeteus; peptides; disulfide-rich peptide (DRP); inhibitory cysteine knot (ICK); venomics; transcriptome; ion channels theraphosidae; Pamphobeteus; peptides; disulfide-rich peptide (DRP); inhibitory cysteine knot (ICK); venomics; transcriptome; ion channels
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Estrada-Gomez, S.; Cardoso, F.C.; Vargas-Muñoz, L.J.; Quintana-Castillo, J.C.; Arenas Gómez, C.M.; Pineda, S.S.; Saldarriaga-Cordoba, M.M. Venomic, Transcriptomic, and Bioactivity Analyses of Pamphobeteus verdolaga Venom Reveal Complex Disulfide-Rich Peptides That Modulate Calcium Channels. Toxins 2019, 11, 496.

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