University of Oulu

Szigetvari, P., Muruganandam, G., Kallio, J., Hallin, E., Fossbakk, A., Loris, R., Kursula, I., Møller, L., Knappskog, P., Kursula, P., Haavik, J. (2018) The quaternary structure of human tyrosine hydroxylase: effects of dystonia‐associated missense variants on oligomeric state and enzyme activity. Journal of Neurochemistry, 148(2), 291-306. doi:10.1111/jnc.14624

The quaternary structure of human tyrosine hydroxylase : effects of dystonia‐associated missense variants on oligomeric state and enzyme activity

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Author: Szigetvari, Peter D.1,2; Muruganandam, Gopinath3,4; Kallio, Juha P.1;
Organizations: 1Department of Biomedicine, University of Bergen, Bergen, Norway
2K.G. Jebsen Centre for Research on Neuropsychiatric Disorders, University of Bergen, Bergen, Norway
3VIB‐VUB Center for Structural Biology, Vlaams Instituut voor Biotechnologie, Brussels, Belgium
4Structural Biology Brussels, Department of Bioengineering Sciences, Vrije Universiteit Brussel, Brussels, Belgium
5Faculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, Finland
6Biocenter Oulu, University of Oulu, Oulu, Finland
7Applied Human Molecular Genetics, Kennedy Center, Department of Clinical Genetics, Copenhagen University Hospital, Rigshospitalet, Glostrup, Denmark
8Department of Clinical Science, University of Bergen, Bergen, Norway
9Center for Medical Genetics and Molecular Medicine, Haukeland University Hospital, Bergen, Norway
10Division of Psychiatry, Haukeland University Hospital, Bergen, Norway
Format: article
Version: published version
Access: open
Online Access: PDF Full Text (PDF, 2.2 MB)
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Language: English
Published: John Wiley & Sons, 2019
Publish Date: 2019-06-20


Tyrosine hydroxylase (TH) is a multi‐domain, homo‐oligomeric enzyme that catalyses the rate‐limiting step of catecholamine neurotransmitter biosynthesis. Missense variants of human TH are associated with a recessive neurometabolic disease with low levels of brain dopamine and noradrenaline, resulting in a variable clinical picture, from progressive brain encephalopathy to adolescent onset DOPA‐responsive dystonia (DRD). We expressed isoform 1 of human TH (hTH1) and its dystonia‐associated missense variants in E. coli, analysed their quaternary structure and thermal stability using size‐exclusion chromatography, circular dichroism, multi‐angle light scattering, transmission electron microscopy, small‐angle X‐ray scattering and assayed hydroxylase activity. Wild‐type (WT) hTH1 was a mixture of enzymatically stable tetramers (85.6%) and octamers (14.4%), with little interconversion between these species. We also observed small amounts of higher order assemblies of long chains of enzyme by transmission electron microscopy. To investigate the role of molecular assemblies in the pathogenesis of DRD, we compared the structure of WT hTH1 with the DRD‐associated variants R410P and D467G that are found in vicinity of the predicted subunit interfaces. In contrast to WT hTH1, R410P and D467G were mixtures of tetrameric and dimeric species. Inspection of the available structures revealed that Arg‐410 and Asp‐467 are important for maintaining the stability and oligomeric structure of TH. Disruption of the normal quaternary enzyme structure by missense variants is a new molecular mechanism that may explain the loss of TH enzymatic activity in DRD. Unstable missense variants could be targets for pharmacological intervention in DRD, aimed to re‐establish the normal oligomeric state of TH.

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Series: Journal of neurochemistry
ISSN: 0022-3042
ISSN-E: 1471-4159
ISSN-L: 0022-3042
Volume: 148
Issue: 2
Pages: 291 - 306
DOI: 10.1111/jnc.14624
Type of Publication: A1 Journal article – refereed
Field of Science: 1182 Biochemistry, cell and molecular biology
Funding: This study was supported by Helse Vest (grant to Jan Haavik); Stiftelsen Kristian Gerhard Jebsen (grant to Jan Haavik), Seventh Framework Programme (grant No. 602805 to Jan Haavik). RL and GM acknowledge the Fonds voor Wetenschappelijk Onderzoek Vlaanderen (grant G0C1213N) and VUB Onderzoeksraad Strategic Research Project (grant SRP‐13) for research support.
Copyright information: © 2018 The Authors. Journal of Neurochemistry published by John Wiley & Sons Ltd on behalf of International Society for Neurochemistry. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.