PhD Scientific Days 2025

Poster Session I. - A: Molecular Medicine

Identification of Interaction Partners of the Human TRESK Background Potassium Channel

Előadó neve

Ms. Debreczeni Dorina

Neptun code

AD6GGT

Előadó munkahelye

Department of Physiology, Semmelweis University

Előadó telefonszáma

+36702579246

Előadó e-mail címe

debreczeni.dorina@stud.semmelweis.hu

Az előadás címe

Identification of Interaction Partners of the Human TRESK Background Potassium Channel

Szerző(k) neve és munkahelye

Dorina Debreczeni¹

1: Department of Physiology, Semmelweis University

Bemutatás módja

Poszter

Szekció

Poster Session I. - A: Molecular Medicine

Language of the presentation

Hungarian

Preferred session

Molecular Medicine

Összefoglaló szövege

Introduction
TRESK (Twik-related spinal cord K⁺ channel) is a unique member of the two-pore domain (K2P) background potassium channel family, expressed in nociceptive primary sensory neurons. Animal studies suggest its role in the pathomechanism of pain and migraine. Previously, we demonstrated that TRESK is regulated by the phosphatase calcineurin via dephosphorylation of serine residues in its intracellular loop, and that its short intracellular C-terminal region also influences channel activity. However, additional mechanisms may also regulate TRESK channel function.
Aims
Our research group has previously conducted in vitro binding assays with various intracellular regions of TRESK, but no such experiments have yet been performed using the full-length channel. The aim of the present study was to identify novel interacting proteins and examine their effects on TRESK channel function.
Method
To search for interacting partners, we used a bait construct of TRESK tagged with an influenza hemagglutinin (HA) epitope and lacking N-glycosylation. Xenopus laevis oocytes were injected with HA-tagged TRESK cRNA or another non-HA-tagged K2P channel as a control. The cells expressed the desired channels in their plasma membranes. The bait protein was pulled down from cell homogenates using anti-HA co-immunoprecipitation, and its presence was verified by anti-HA immunoblotting. Eluates in SDS sample buffer were analyzed by LC–MS/MS.
Results
Protein identification revealed two promising candidates. One is a kinase (9 unique peptides, 21% coverage), with a potential binding motif in the intracellular loop. The other is a protein involved in regulating intracellular vesicle transport (5 unique peptides, 8% coverage). We cloned the cDNAs of both putative interaction partners from HEK293T cells. Co-expression with TRESK in Xenopus oocytes, followed by two-electrode voltage-clamp recordings, demonstrated their effects on channel function.
Conclusion
We identified two novel interaction partners of the human TRESK potassium channel. These findings enhance understanding of TRESK regulation and may help elucidate its role in nociceptive signaling and pain modulation.
Funding
Supported by the 2024-2.1.1-EKÖP-2024-00004 University Research Scholarship Programme of the Ministry for Culture and Innovation from the source of the National Research, Development and Innovation Fund.

University

Semmelweis University

Supervisor

Gábor Czirják, MD., PhD., DSc.

Publication of my abstract

I do not give consent to the publication of my abstract on the website of the congress.

phd.section.field

before finishing undergraduate studies (TDK, MD-PhD)

Kind

Szabad

Status

elfogadva

Accepted presentation method

poszter

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

9053

Start

17:30

End

17:36