Biological Information

Background Information:

Serotonin 5-HT receptors belong to the superfamily of G protein-coupled seven transmembrane proteins with the exception of the 5-HT3 subtype, which belongs to the ligand gated cation superfamily of receptors. G-protein-coupled receptors constitute one of the major signal transduction systems in eukaryotic cells. Coding sequences for these receptors, in those regions believed to contribute to the agonist-antagonist binding site, are strongly conserved across mammalian species. Serotonin 5-HT5A receptors are widely present in the central nervous system but not or minimally in peripheral tissues. The 5-HT5A receptor class couples preferentially to GS to promote cAMP formation. Serotonin 5-HT5A receptor agonism may be hallucinatory and alter circadian rhythms.

No. of Assays on Panel:

2

Assays on Panel:

Family

GPCR

Name

Item

355400-0

Protein

5-HT5A

Organism

Human

Assay Sub Type

Cell Based

Family

GPCR

Name

Item

355400-1

Protein

5-HT5A

Organism

Human

Assay Sub Type

Cell Based

Family:

GPCR

Sub Family:

Serotonin

Class:

Class A

Protein Name:

5-HT5A

Uniprot Number:

P47898

Protein Aliases:

5-HT-5A|5-HT5A|Serotonin receptor 5A

Gene Name:

HTR5A

Gene ID:

3361

Gene Aliases:

5-HT5A

Accession Number:

NM_024012

Organism:

Human

Construct Details:

Full Length, Recombinant

Cell Type:

CHO-K1

Assay Information

Assay Type:

Functional

Assay Sub Type:

Cell Based

Functional Mode:

Agonist & Antagonist

Detection Method:

Spectrofluorimetry

Measured Response:

Fluorescence

Testing Information

Procedure Summary:

Human recombinant serotonin 5-HT5A receptor stably expressed in CHO-K1 cells are used. Test compound and/or vehicle is incubated with the cells (5 x 10^5 /ml) in incubation buffer of fluo-4 direct calcium kit for 20 minutes at RT. Test compound-induced increase of fluorescence by 50 percent or more (≥50%) relative to the 10 μM 5-CT response indicates possible serotonin 5-HT5A receptor agonist activity. Test compound-induced inhibition of 0.03 μM 5-CT-induced fluorescence response by 50 percent or more (≥50%) indicates receptor antagonist activity.

Incubation:

20 min at RT

Control Inhibitor:

Methiothepin

Control Activator:

5-Carboxamidotryptamine

Criteria For Significance:

≥ 50% increase in fluorescence relative to 5-CT response, ≥ 50% inhibition in fluorescence relative to 5-CT-induced response

Test Concentration / Dose:

Compounds are screened at 10, 3, 1, 0.3, 0.1, 0.03, 0.01 and 0.003 μM

Test Sample Requirements:

75 μl of 10 mM stock or 1 mg (pre-weighed) powder (assuming FW ≦500 and purity 100%)

Minimum Order Quantity:

16

Turnaround Time

Standard:

20 Business Days

Reference Compound Data

PubChem ID

1809

Name

5-Carboxamidotryptamine

Measurement

0.0093μM - EC50

PubChem ID

4106

Name

Methiothepin

Measurement

0.0064μM - IC50

PubChem ID

9681

Name

Methysergide

Measurement

0.42μM - IC50

PubChem ID

11168182

Name

SB-699,551

Measurement

2.3μM - IC50

PubChem ID

Name

Serotonin (5-HT)

Measurement

0.028μM - EC50

Clinical Relevance

Therapeutic Area:

CNS/Addiction/Pain

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei

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