Sudeep Karki, Mirko M. Maksimainen, Lari Lehtiö, Tommi Kajander, Inhibitor screening assay for neurexin-LRRTM adhesion protein interaction involved in synaptic maintenance and neurological disorders, Analytical Biochemistry, Volume 587, 2019, 113463, ISSN 0003-2697, https://doi.org/10.1016/j.ab.2019.113463
Inhibitor screening assay for neurexin-LRRTM adhesion protein interaction involved in synaptic maintenance and neurological disorders
|Author:||Karki, Sudeep1; Maksimainen, Mirko M.2; Lehtiö, Lari2;|
1Institute of Biotechnology, University of Helsinki, Finland
2Faculty of Biochemistry and Molecular Medicine and Biocenter Oulu University of Oulu, Finland
|Persistent link:|| http://urn.fi/urn:nbn:fi-fe202002044535
|Publish Date:|| 2021-09-28
Synaptic adhesion molecules, including presynaptic neurexins (NRXNs) and post-synaptic leucine-rich repeat transmembrane (LRRTM) proteins are important for development and maintenance of brain neuronal networks. NRXNs are probably the best characterized synaptic adhesion molecules, and one of the major presynaptic organizer proteins. The LRRTMs were found as ligands for NRXNs. Many of the synaptic adhesion proteins have been linked to neurological cognitive disorders, such as schizophrenia and autism spectrum disorders, making them targets of interest for both biological studies, and towards drug development. Therefore, we decided to develop a screening method to target the adhesion proteins, here the LRRTM-NRXN interaction, to find small molecule probes for further studies in cellular settings. To our knowledge, no potent small molecule compounds against the neuronal synaptic adhesion proteins are available. We utilized the AlphaScreen technology, and developed an assay targeting the NRXN-LRRTM2 interaction. We carried out screening of 2000 compounds and identified hits with moderate IC₅₀-values. We also established an orthogonal in-cell Western blot assay to validate hits. This paves way for future development of specific high affinity compounds by further high throughput screening of larger compound libraries using the methods established here. The method could also be applied to screening other NRXN-ligand interactions.
|Type of Publication:||
A1 Journal article – refereed
|Field of Science:||
1182 Biochemistry, cell and molecular biology
This work was funded by Jane and Aatos Erkko Foundation (TK), and Academy of Finland (grant no. 287063 and 294085 for LL).
|Academy of Finland Grant Number:||
287063 (Academy of Finland Funding decision)
294085 (Academy of Finland Funding decision)
© 2019. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/.