Tyrosine Hydroxylase Rat Enzymatic LeadHunter Assay - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Tyrosine hydroxylase (tyrosine-3-monooxygenase, tyrosine oxidase, E.E.1.14.13.41) which catalyzes the hydroxylation of tyrosine is generally regarded as a rate-limiting step in the biosynthesis of catecholamines. The enzyme is activated following stimulation of adrenergic nerves or the adrenal medulla. Tyrosine hydroxylase is a substrate for cyclic AMP-dependent kinase, and kinase-catalyzed phosphorylation is associated with increased hydroxylase activity. The enzyme is also a substrate for Ca2+-activated protein kinases. In addition, tyrosine hydroxylase is inhibited by catechol compounds in a manner that is competitive with its pterin cofactor, tetrahydrobiopterin. It is thus subject to end-producr feedback inhibition. A potent and selective inhibitor of this enzyme may prove useful as a therapeutic agent in the treatment of hypertension as well as schizophrenia and tardive dyskinesia.
Family:
Neurotransmitter Metabolism
Sub Family:
Amino Acid Hydroxylase
Protein Name:
Tyrosine Hydroxylase
Uniprot Number:
P07101
Protein Aliases:
tyrosine 3-monooxygenase
Gene Name:
TH
Gene ID:
7054
Gene Aliases:
DYT5b
Accession Number:
NM_012740
Organism:
Rat
Construct Details:
Endogenous
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Radiometry
Measured Response:
Scintillation
Testing Information
Procedure Summary:
Rat brain tyrosine hydroxylase is used. Test compound and/or vehicle is preincubated with 4.9 mg/ml enzyme‡ in modified MES buffer pH 6.0 for 15 minutes at 37⁰C. The reaction is initiated by addition of 10 μM L-tyrosine and 0.5 μCi L-[3,5-3H]Tyrosine for another 20 minute incubation period and terminated by further addition of 1 N HCl. An aliquot is removed and counted to determine the amount of [3H]H2O formed. Compounds are screened at 10 μM.
Substrate:
10 µM L-[3, 5-³H]Tyrosine + L-Tyrosine
Incubation:
20 min at 37°C
Control Inhibitor:
Alpha-Methyl-L-P-Tyrosine
Criteria For Significance:
≥ 50% of max stimulation or inhibition
Test Concentration / Dose:
Suggested concentration for initial testing is 10 uM
Test Sample Requirements:
Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).
Minimum Order Quantity:
2
Turnaround Time
Standard:
15 Business Days
Reference Compound Data
Clinical Relevance
Therapeutic Area:
CNS/Addiction/Pain
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
Questions? Contact Us
You can easily get in touch with a Eurofins Discovery Expert or get help from our Customer Service team. Reach out with questions, or to discuss your specific discovery challenges.
