Experimental and Computational Analysis of Newly Identified Pathogenic Mutations in the Creatine Transporter SLC6A8.

Autor: Ferrada E; CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria. Electronic address: EFerrada@cemm.oeaw.ac.at., Wiedmer T; CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria., Wang WA; CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria., Frommelt F; CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria., Steurer B; CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria., Klimek C; CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria., Lindinger S; CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria., Osthushenrich T; Bayer AG, Pharmaceuticals, Wuppertal, Germany., Garofoli A; CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria., Brocchetti S; Axxam SpA, Bresso, Milano, Italy., Bradberry S; Axxam SpA, Bresso, Milano, Italy., Huang J; Department of Pharmaceutical Sciences, University of Vienna, Vienna, Austria., MacNamara A; Bayer AG, Pharmaceuticals, Wuppertal, Germany., Scarabottolo L; Axxam SpA, Bresso, Milano, Italy., Ecker GF; Department of Pharmaceutical Sciences, University of Vienna, Vienna, Austria., Malarstig A; Pfizer Worldwide Research, Development and Medical, Stockholm, Sweden., Superti-Furga G; CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria; Center for Physiology and Pharmacology, Medical University of Vienna, Vienna, Austria. Electronic address: GSuperti@cemm.oeaw.ac.at.
Jazyk: angličtina
Zdroj: Journal of molecular biology [J Mol Biol] 2024 Jan 15; Vol. 436 (2), pp. 168383. Date of Electronic Publication: 2023 Dec 07.
DOI: 10.1016/j.jmb.2023.168383
Abstrakt: Creatine is an essential metabolite for the storage and rapid supply of energy in muscle and nerve cells. In humans, impaired metabolism, transport, and distribution of creatine throughout tissues can cause varying forms of mental disability, also known as creatine deficiency syndrome (CDS). So far, 80 mutations in the creatine transporter (SLC6A8) have been associated to CDS. To better understand the effect of human genetic variants on the physiology of SLC6A8 and their possible impact on CDS, we studied 30 missense variants including 15 variants of unknown significance, two of which are reported here for the first time. We expressed these variants in HEK293 cells and explored their subcellular localization and transport activity. We also applied computational methods to predict variant effect and estimate site-specific changes in thermodynamic stability. To explore variants that might have a differential effect on the transporter's conformers along the transport cycle, we constructed homology models of the inward facing, and outward facing conformations. In addition, we used mass-spectrometry to study proteins that interact with wild type SLC6A8 and five selected variants in HEK293 cells. In silico models of the protein complexes revealed how two variants impact the interaction interface of SLC6A8 with other proteins and how pathogenic variants lead to an enrichment of ER protein partners. Overall, our integrated analysis disambiguates the pathogenicity of 15 variants of unknown significance revealing diverse mechanisms of pathogenicity, including two previously unreported variants obtained from patients suffering from the creatine deficiency syndrome.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2023 CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences GmbH. Published by Elsevier Ltd.. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: