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

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 ma...

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Bibliographic Details
Published in:Toxins 2019-08, Vol.11 (9), p.496
Main Authors: Estrada-Gomez, Sebastian, Cardoso, Fernanda Caldas, Vargas-Muñoz, Leidy Johana, Quintana-Castillo, Juan Carlos, Arenas Gómez, Claudia Marcela, Pineda, Sandy Steffany, Saldarriaga-Cordoba, Monica Maria
Format: Article
Language:English
Subjects:
Biological activity
Calcium
Calcium channels
Calcium channels (voltage-gated)
Calcium influx
disulfide-rich peptide (DRP)
Fractions
High performance liquid chromatography
inhibitory cysteine knot (ICK)
Ion channels
Ions
Liquid chromatography
Mass spectrometry
Mass spectroscopy
Molecular weight
Neurotoxicity
Pamphobeteus
Peptides
Pipelines
Polyamines
Potassium
Proteins
Scientific imaging
Sodium
Sodium channels (voltage-gated)
theraphosidae
Toxins
Transcription
transcriptome
Transcriptomes
Venom
venomics
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Summary: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 venom showed activity on Ca 2.2, Ca 3.2, and Na 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 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 venom is an important source of disulfide-rich proteins expressing at least eight superfamilies.
ISSN:2072-6651
2072-6651
DOI:10.3390/toxins11090496