Biophysical study of the interaction between TRPM3 and the blocking compound. - Belgium
KU Leuven (company)
Chronicpain is a major health problem affecting millions of people worldwide. Abouthalf of people with chronic pain report inadequate pain control. Despite decades of research, chronic painremains poorly understood and notoriously hard to control. Therefore, there isa high need for the identification of potential drug targets as a startingpoint in the development of novel painkillers. Several members of the TransientReceptor Potential (TRP) ion channel superfamily are highly expressed insensory neurons and are involved in the detection of painful stimuli. These nociceptiveTRP channels have been proposed and investigated as potential new drug targets. Recently, we have identified TRPM3 asan important pain sensor highly expressed in the plasma membrane of sensoryneurons. Interestingly activation of TRPM3 by neurosteroids are noxious heatinduces pain. Therefore, the TRPM3 protein is a promising target for thetreatment of chronic pain.
Tounderstand the (patho)physiological effects of endogenous ligands, to predictthe effects of exogenous ligands, and to rationally design drug-basedtherapeutic approaches, it is crucial to have detailed insights into themolecular and biophysical mechanisms of channel-ligand interactions. Here, weaim to determine ligand-interaction sites, perform structure-activity relationanalysis, and to elucidate the precise biophysical consequences of compoundbinding on channel gating.
Methodology: Patch-clamprecordings in HEK293 cells expressing human TRPM3, in either its wild type ormutated form and in sensory neurons of the somatosensory system.
TRPM3is voltage-gated, and by measuring whole-cell currents during voltage-steps,both in the absence and the presence of different concentrations of the leadcompounds, we can determine the effects of TRPM3 antagonists on the channels’ voltagedependence and gating kinetics.
It iscurrently unknown whether TRPM3 antagonists compete with binding of endogenousneurosteroid agonists such as pregnenolone sulphate (PS), or where and how theyinteract with the channel. To identify the interaction sites of ligands,we will compare the kinetics of ligand action in different modes of thepatch-clamp technique (whole-cell, inside-out, outside-out and cell-attached).This will allow us to determine whether the compounds act on the intra- orextracellular domains of the channel or rather at the level of thetransmembrane domains. Second, mutations will be introduced in selected regionsof the channel, and the consequences on ligand-channel interaction will be analyzedas outlined above. Mutagenesis will be focused on channel regions that areidentified based on preliminary data and/or bioinformatics approaches.
You have a
- Master degree in Biology, Biomedical Sciences or Bio-engineer
- You obtained good grades
- Interested in neurobiology and electrophysiology
- Very good writing and language skills in English
- Very good communication and organization skills. You are a team player who can work together with researchers
- Able to work independently
- a fulltime employment for one year, which can be extended to 4 years, with the opportunity to obtain a PhD degree.
- a stimulating work and learning environment in which you acquire expertiseas a multidisciplinary researcher
- Starting date is 1 September 2017 (to be discussed)