Magic™ In Vitro Cell-based Glycoprotein Hormone Receptor Functional Assay Service

The Importance of Glycoprotein Hormone Receptors

Glycoprotein hormone receptors (GPHRs) are rhodopsin-like GPCRs with a substantial extracellular N-terminal region that is essential for hormone recognition and binding. TSH receptor (Thyrotropin), FSH receptor (Follitropin), and LH receptor (Lutropin) are three types of GPHRs. Although their major physiological function is to support gonadal progress, several studies have revealed that mutations in these receptors cause the emergence of certain disorders that are difficult to treat. Whereupon, there is an urgent need for an in-depth interpretation of GPHRs.

Fig.1 Model of the TSH receptor.¹

Our Magic™ In Vitro Cell-Based Glycoprotein Hormone Receptor Functional Assay Service

To boost the development of drug discovery, Creative Biolabs provides a simple and effective Magic™ in vitro cell-based glycoprotein hormone receptor functional assay service for global customers.

In our Magic™ in vitro cell-based glycoprotein hormone receptor functional assay service, we integrate in vitro cell-based assay with high sensitivity and effectiveness to glycoprotein hormone receptors functional assay. Through investigating changes in second messenger concentrations and downstream calcium fluxes resulting from receptor activation, this assay allows us to identify compounds and their ability to modify glycoprotein hormone receptors. Together with our high-quality comprehensive customized services, Creative Biolabs' Magic™ in vitro cell-based glycoprotein hormone receptor functional assay service can satisfy the diverse demands of global customers.

Fig.2 Signaling pathways downstream of LHCGR.²

Benefit for You

Our Magic™ in vitro cell-based glycoprotein hormone receptor functional assay service can benefit you with:

Fig.3 Benefit of our Magic™ in vitro cell-based glycoprotein hormone receptor functional assay service.

Case Study

Representative Data 1: Key signaling pathways in HEK-293 cells recombinantly expressing LHCGR (HEK-LHCGR) were examined to confirm gonadotropin bioactivity. Cells were challenged with either hCG or LH and compared to wild-type HEK-293 cells (HEK-WT) in parallel studies.

• cAMP assay: Intracellular cAMP levels increased significantly in HEK-LHCGR cells after 5 minutes of exposure to either hCG or LH. Inductions were concentration-dependent, with pEC50 values for hCG of 9.671 (213 pM) and LH of 9.370 (426 pM), with peak responses at 10 nM. HEK-WT cells, on the other hand, did not react.

Fig.4 cAMP responses to hCG and LH in HEK-WT and HEK-LHCGR cells.²

• Ca²⁺ assay: Calbryte-520, a Ca²⁺-sensitive dye, was also added to HEK cells before they were challenged with high doses of hCG or LH (1 M). In HEK-LHCGR cells, both agonists induced strong Ca²⁺ responses, but not in HEK-WT cells.

Fig.5 Ca²⁺ signals in HEK-WT and -LHCGR cells after hCG and LH treatment, respectively.²

• ERK phosphorylation assay: Both ligands resulted in a significant and prolonged elevation of phosphorylated (active)-ERK in HEK-LHCGR cells, but HEK-WT cells did not react.

Fig.6 The effects of hCG and LH on phospho-ERK levels in wild-type and LHCGR-expressing HEK cells.²

Representative Data 2: The effect of FSHR (271-275) peptide on FSH binding to FSHR was investigated by incubating the peptide with HEK-rFshr cell membrane preparation in the presence of radiolabelled hFSH. These findings showed that FSHR bound to FSHR (271-275) peptide revealed enhanced FSH binding and activity when compared to FSHR without peptide.

Fig.7 Effect of FSHR (271-275) peptide on FSH-FSHR binding and FSH-induced cAMP production.³

As an industry leader in GPCR functional assays, Creative Biolabs is dedicated to providing every customer with the most advanced techniques as well as top-ranking services. For more details about our Magic™ in vitro cell-based glycoprotein hormone receptor functional assay service, please don't hesitate to get in touch with us. All of us here are eager to get to work with you right away.

References

1. Galofré, Juan C., et al. "Targeting thyroid diseases with TSH receptor analogs." Endocrinología y Nutrición 60.10 (2013): 590-598.
2. Mann, O. N., et al. "Expression and function of the luteinizing hormone choriogonadotropin receptor in human endometrial stromal cells." Scientific Reports 12.1 (2022): 8624.
3. Prabhudesai, Kaushiki S., et al. "A 5-mer peptide derived from hinge region of hFSHR can function as a positive allosteric modulator in vivo." Biochimica et Biophysica Acta (BBA)-Biomembranes 1863.1 (2021): 183492.

For Research Use Only.