Molecular mechanism for pterin-mediated inactivation of tyrosine hydroxylase: Formation of insoluble aggregates of tyrosine hydroxylase
Abstract
Tyrosine hydroxylase (TH), an iron-containing enzyme, catalyzes the first and rate-limiting step of catecholamine biosynthesis, and requires tetrahydrobiopterin (BH4) as a cofactor. We found that preincubation of recombinant human TH with BH4 results in the irreversible inactivation of the enzyme at a concentration far less than the Km value toward BH4 in spite of its cofactor role, whereas oxidized biopterin, which has no cofactor activity, does not affect the enzyme activity. We show that TH is inactivated by BH4 in competition with the binding of dopamine. The sequential addition of BH4 to TH results in a gradual decrease in the intensity of the fluorescence and CD spectra without changing their overall profiles. Sedimentation velocity analysis demonstrated an association of TH molecules with each other in the presence of BH4, and studies using gel-permeation chromatography, turbidity measurements, and transmission electron microscopy demonstrated the formation of amorphous aggregates with large molecular weights following the association of the TH proteins. These results suggest that BH4 not only acts as a cofactor, but also accelerates the aggregation of TH. We propose a novel mechanism for regulating the amount of TH protein, and discuss its physiological significance.
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Recombinant human TH protein forms insoluble aggregates in the presence of tetrahydrobiopterin in vitro.
“Urano et al. reported that recombinant human TH protein forms insoluble aggregates in the presence of tetrahydrobiopterin in vitro [99].”
From Degradation of Tyrosine Hydroxylase by the Ubiquitin-Proteasome System in the Pathogenesis of Parkinson’s Disease and Dopa-Responsive Dystonia (2020) · cites this paper as [99]
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- Authors
- Urano, F., Hayashi, N., Arisaka, F., Kurita, H., Murata, S., & Ichinose, H.
- Affiliations
- Tokyo Institute of Technology (JP)
- Corresponding author
- Urano, Fumi
- Venue
- The Journal of Biochemistry
- Publisher
- Oxford University Press (OUP)
- Published
- 2006-04-01
- Volume
- 139
- Issue
- 4
- Pages
- 625-635
- ISSN
- 0021-924X (ISSN-L)
- 1756-2651
- OpenAlex
- W2065461666
- Cited by
- 30
- References
- 40
- Citations by year
- 2025: 1 · 2024: 1 · 2023: 1 · 2021: 2 · 2020: 1 · 2018: 3 · 2017: 1 · 2015: 2 · 2013: 4
- FWCI
- 2.05
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- The Journal of Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institute of Diabetes and Digestive and Kidney Diseases (R01DK067493)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Clinical Biochemistry › Metabolism and Genetic Disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Amino Acid Enzymes and Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
- Keywords
- tyrosine hydroxylase, tetrahydrobiopterin, enzyme inactivation, protein aggregation, catecholamine synthesis, dopamine binding, cofactors, iron-containing enzymes
- MeSH terms
- Biopterins, Centrifugation, Density Gradient, Chromatography, Gel, Circular Dichroism, Dopamine, Electrophoresis, Polyacrylamide Gel, Enzyme Activation, Humans, Kinetics, Models, Chemical, Pterins, Recombinant Proteins, Spectrometry, Fluorescence, Tyrosine 3-Monooxygenase, Microscopy, Electron, Transmission
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