title: Margatoxin-bound quantum dots as a novel inhibitor of the voltage-gated ion channel Kv1.3.
abstract: Venom-derived ion channel inhibitors have strong channel selectivity, potency, and stability; however, tracking delivery to their target can be challenging. Herein, we utilized luminescent quantum dots (QDs) conjugated to margatoxin (MgTx) as a traceable vehicle to target a voltage-dependent potassium channel, Kv1.3, which has a select distribution and well-characterized role in immunity, glucose metabolism, and sensory ability. We screened both unconjugated (MgTx) and conjugated MgTx (QD-MgTx) for their ability to inhibit Shaker channels Kv1.1 to Kv1.7 using patch-clamp electrophysiology in HEK293 cells. Our data indicate that MgTx inhibits 79% of the outward current in Kv1.3-transfected cells and that the QD-MgTx conjugate is able to achieve a similar level of block, albeit a slightly reduced efficacy (66%) and at a slower time course (50% block by 10.9 ± 1.1 min, MgTx; vs. 15.3 ± 1.2 min, QD-MgTx). Like the unbound peptide, the QD-MgTx conjugate inhibits both Kv1.3 and Kv1.2 at a 1 nM concentration, whereas it does not inhibit other screened Shaker channels. We tested the ability of QD-MgTx to inhibit native Kv1.3 expressed in the mouse olfactory bulb (OB). In brain slices of the OB, the conjugate acted similarly to MgTx to inhibit Kv1.3, causing an increased action potential firing frequency attributed to decreased intraburst duration rather than interspike interval. Our data demonstrate a retention of known biophysical properties associated with block of the vestibule of Kv1.3 by QD-MgTx conjugate compared to that of MgTx, inferring QDs could provide a useful tool to deliver ion channel inhibitors to targeted tissues in vivo.
authors.affiliation: Institute of Molecular Biophysics, Florida State University, Tallahassee, Florida, USA.
authors.forename: Austin B
authors.initials: AB
authors.lastname: Schwartz
authors.affiliation: Department of Chemistry and Biochemistry, Florida State University, Tallahassee, Florida, USA.
authors.forename: Anshika
authors.initials: A
authors.lastname: Kapur
authors.affiliation: Department of Chemistry and Biochemistry, Florida State University, Tallahassee, Florida, USA.
authors.forename: Wentao
authors.initials: W
authors.lastname: Wang
authors.affiliation: Program in Neuroscience, Florida State University, Tallahassee, Florida, USA.
authors.forename: Zhenbo
authors.initials: Z
authors.lastname: Huang
authors.affiliation: Program in Neuroscience, Florida State University, Tallahassee, Florida, USA.|Department of Biological Science, Florida State University, Tallahassee, Florida, USA.
authors.forename: Erminia
authors.initials: E
authors.lastname: Fardone
authors.affiliation: Department of Chemistry and Biochemistry, Florida State University, Tallahassee, Florida, USA.
authors.forename: Goutam
authors.initials: G
authors.lastname: Palui
authors.affiliation: Department of Chemistry and Biochemistry, Florida State University, Tallahassee, Florida, USA.
authors.forename: Hedi
authors.initials: H
authors.lastname: Mattoussi
authors.affiliation: Institute of Molecular Biophysics, Florida State University, Tallahassee, Florida, USA.|Program in Neuroscience, Florida State University, Tallahassee, Florida, USA.|Department of Biological Science, Florida State University, Tallahassee, Florida, USA.
authors.forename: Debra Ann
authors.initials: DA
authors.lastname: Fadool
mesh_terms: D000200:Action Potentials / Q000187:drug effects / Q000502:physiology; D000818:Animals; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D051664:Kv1.3 Potassium Channel / Q000037:antagonists & inhibitors* / Q000502:physiology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009498:Neurotoxins / Q000378:metabolism / Q000494:pharmacology*; D009830:Olfactory Bulb / Q000187:drug effects / Q000502:physiology; D045663:Quantum Dots / Q000008:administration & dosage* / Q000378:metabolism; D012604:Scorpion Venoms / Q000378:metabolism / Q000494:pharmacology
chemical_list: D051664:Kv1.3 Potassium Channel; D009498:Neurotoxins; D012604:Scorpion Venoms; C077910:margatoxin
keywords: K channel; MgTx; nanoparticle; patch clamp; pore blocker; potassium channel