Thyroid Stimulating Hormone Receptor (TSHR) Structure and Function

Introduction

Figure 1: TSHR binds TSH in the HPT signaling axis pathway, which regulates metabolism and growth.

In humans, the hypothalamic-pituitary-thyroid (HPT) signaling axis regulates functions including metabolism, growth, organ development, and neural differentiation ^[1]. In this pathway, the thyroid stimulating hormone receptor (TSHR) activates transcription of thyroid hormones in response to ligand binding by thyroid stimulating hormone (TSH). After a brief introduction to the biological significance of TSHR, this page explores the structure of TSHR and its significance to TSH binding and receptor activation.

Biological Significance of TSHR

The HPT signaling axis involves the brain, thyroid gland, and bloodstream circulation. In the first step of the pathway, thyrotropin releasing hormone (TRH) is secreted by the hypothalamus, which in turn stimulates the anterior pituitary gland to produce TSH ^[1]. TSH binds to TSHR on the surface of thyroid cells and triggers the production of thyroid hormones thyroxine (T3) and triiodothyronine (T4) through G-protein coupled receptor (GPCR) signaling ^[2]. T3 and T4 circulate in the bloodstream and enter cells via thyroid hormone transporters to regulate metabolic functions. Additionally, T3 and T4 act in a negative feedback loop to inhibit further TSH production ^[1].

Dysregulation of TSHR can lead to disease. In Grave's disease, antibody analogs of TSH cause overactivation of TSHR, leading to clinical symptoms of hyperthyroidism ^[2]. In contrast, congenital mutations which inactivate TSHR can lead to hypothyroidism, which results in growth retardation and neurologic impairment if left untreated ^[1].

Structural Overview of TSHR

There are three main domains of the Thyroid Stimulating Hormone Receptor. First is the shown in green. Region is concave in shape and is made up of primarily beta sheets and rich in Leucine. It is also called the leucine rich region ^[3]. This is also the domain for key Lysine residues, which play a key role in binding. Second is the shown in pink. This domain is made up of seven helices and undergoes a conformation change upon ligand binding that activates the GPCR signal cascade (GPCR is shown in yellow) ^[4]. The third region of the TSHR is the shown in orange. The Hinge Region plays a key role in the movement and stability of the TSHR.

TSHR Activation

Hinge Motion

Central to the biological function of TSHR is its hinge motion which allows for transition between active and inactive states. Deformation of the hinge region accommodates up-and-down rotation of the extracellular domain as a rigid body about an imaginary 55 degree axis. When the extracellular domain is upright, the receptor actively signals for thyroid hormone production. When the extracellular domain is hinged down, the receptor is inactive and no signaling activation occurs. Notably, transition between the two states occurs spontaneously; favoring of the active or inactive conformation is influenced by hinge interactions and ligand binding ^[5].

At the following link is an animation of TSHR transition between the two states: Image:TSHR MorphBetterAngle.mp4. The following trends can be observed in the video or viewed through an in the molecule viewer:

1. Slinky-like deformation of the hinge region shifts the region 5 Angstroms over the course of the movement ^[5].
2. Stretching of the hinge pulls on the linked helices in the transmembrane domain, shifting about 4 Angstroms inward ^[5].

Stabilizing Interactions in the Hinge

To understand stabilizing interactions which accommodate the hinge motion, the hinge region can be subdivided into the , which lies at the intersection of the extracellular and transmembrane domains; , which sticks up and serves as a binding platform for the TSH ligand; the region, which connects helix 1 with the p10 region; and the region, a conserved 10-amino acid sequence which connects to transmembrane helix 7 and undergoes most of the deformation ^{[5] [4]}.

Two key disulfide bridges within the hinge region help to maintain its structure and orientation ^[4].

1. The connects the hinge helix with the linker region
2. The connects the hinge helix with the p10 region.

The upright, active conformation of the hinge is stabilized by its respective interactions wit the EC and TM domains ^[5].

1. A occurs between Y279 in the hinge helix and I486 in EC loop region 1, which protrudes from the TM helices.
2. An occurs between K660 in TM helix 7 and E409 in the p10 region.

If the stabilizing interactions are disrupted, TSHR function is affected. For instance, the mutation I496F has been observed to cause constitutive receptor activation and decreased sensitivity to the TSH ligand, suggesting that the bulkier phenylalanine strengthens the hydrophobic interaction too much leading to overactivation. Contrastingly, TSHR underactivation results from disrupting the ionic interaction with an E409A mutation, which is associated with diminished receptor activation and TSH potency ^[5].

Ligand Binding

Binding of Thyroid Stimulating Hormone to TSHR

The Thyroid Stimulating Hormone by complementary shape. The ECD and is curved and compliments the curvature of TSH similar to how a baseball fits into a glove. There are also several key ionic interactions between the TSH and TSHR. The key ionic interactions occurs in the which is highlighted in yellow. The seatbelt region is located in the beta subunit of the TSH. is Glu118 from TSH and Lys58 from the ECD. is between Asp111 from the TSH and Lys209 from the ECD. These interactions form salt bridges between the ECD and the TSH which allows for specificity of binding for TSH to TSHR ^[4],^[5].

Other key interactions that allow for specificity of binding are between the helix 1. Helix 1 contains several polar residues that interact with surrounding nonpolar residues like Leu62, Arg54, and Phe17. These interactions increase the activation potency and helps activate the push and pull mechanism of the hinge region ^[4],^[5].

Blocking TSHR in Active/Inactive States

TSHR in active and inactive binding states. Left is TSH bound to TSHR. Middle is M22 bound to TSHR. Right is CS-17 bound to TSHR.

The interactions between the ligand and the receptor have important consequences for disease states. In the image shown to the right are three different states of TSHR. The right-most structure is TSH bound to TSHR and is in it's upright active conformation. Comparing this is M22, shown in the middle, TSH binding offer not steric hinderance to keep it from being released. M22 can manifest with elevated levels of thyroid hormones which could be found in a person with Graves disease. M22 bound to TSHR is in the upright state and prevents transition to the down state because of the steric clash with the membrane which causes constitutive activation and symptoms of Graves disease. On the right most side is CS-17 bound to the TSHR. In contrast to TSH and M22 binding, CS-17 binds and locks TSHR in the down, inactive conformation. This prevents the signaling cascade to translation and causes constitutive inactivation ^[5].

These different ways to active and inactive TSHR could represent potential therapies for someone with graves disease or other thyroid related diseases with overacting TSH binding. Whereas current therapies target T3/T4 synthesis or destroy the gland using artificial hormones, these diseases could instead be targeted with something like M22 or CS-17.

References

1. ↑ ^{1.0 1.1 1.2 1.3} Brent GA. Mechanisms of thyroid hormone action. J Clin Invest. 2012;122(9):3035-3043. DOI: 10.1172/JCI60047
2. ↑ ^{2.0 2.1} Chu YD, Yeh CT. The Molecular Function and Clinical Role of Thyroid Stimulating Hormone Receptor in Cancer Cells. Cells. 2020;9(7):1730. DOI:10.3390/cells9071730
3. ↑ Kleinau G, Worth CL, Kreuchwig A, et al. Structural–Functional Features of the Thyrotropin Receptor: A Class A G-Protein-Coupled Receptor at Work. Frontiers in Endocrinology. 2017;8. Accessed April 2, 2023. DOI: 10.3389/fendo.2017.00086
4. ↑ ^{4.0 4.1 4.2 4.3 4.4} Duan J, Xu P, Luan X, Ji Y, He X, Song N, Yuan Q, Jin Y, Cheng X, Jiang H, Zheng J, Zhang S, Jiang Y, Xu HE. Hormone- and antibody-mediated activation of the thyrotropin receptor. Nature. 2022 Aug 8. pii: 10.1038/s41586-022-05173-3. doi:, 10.1038/s41586-022-05173-3. PMID:35940204 doi:http://dx.doi.org/10.1038/s41586-022-05173-3
5. ↑ ^{5.0 5.1 5.2 5.3 5.4 5.5 5.6 5.7 5.8} Faust B, Billesbolle CB, Suomivuori CM, Singh I, Zhang K, Hoppe N, Pinto AFM, Diedrich JK, Muftuoglu Y, Szkudlinski MW, Saghatelian A, Dror RO, Cheng Y, Manglik A. Autoantibody mimicry of hormone action at the thyrotropin receptor. Nature. 2022 Aug 8. pii: 10.1038/s41586-022-05159-1. doi:, 10.1038/s41586-022-05159-1. PMID:35940205 doi:http://dx.doi.org/10.1038/s41586-022-05159-1