Claus Juul Løland Group – University of Copenhagen

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Neuropharmacology > Research > Claus Juul Løland Group

Research

The main focus of the LolandLab is to investigate the structure-function relationships in neurotransmitter:sodium symporters (NSS) with the aim of elucidating the molecular mechanisms behind substrate transport. The molecular basis behind transport inhibition by e.g. antidepressants and illicit drugs such as cocaine is investigated as well as the identification and characterization of novel inhibitor binding sites. To address these issues we have employed two general approaches. The first approach is molecular pharmacology using classical site-directed mutagenesis and radioligand binding/uptake assays to probe for novel binding sites and to determine binding poses and –conformations of both well-known drugs (cocaine, amphetamine, antidepressants) as well as novel inhibitors with the aim of developing a medical treatment against cocaine addiction. These assays are performed on cell cultures expressing the NSS protein of interest. The other approach is the use of advanced biophysical methods on purified protein in direct assessment of conformational changes. This includes fluorescence spectroscopy and -life-time techniques on detergent solubilized transporters with site-selectively incorporated fluorescent probes. Here we use transition metal-ion FRET (tmFRET) or environmental sensitive dyes to monitor conformational changes induced by the binding of substrates, ions or mutations in conformational important regions (gating domains). Together with collaborators, we also have employed Hydrogen-Deuterium exchange mass spectrometry (HDx-MS), protein crystallography, neutron scattering, solid state NMR etc.

Quencing of fluorescin emission coubled to NSS protein by Ni2+