| 2011 |
GPR133 (ADGRD1) couples to Gs protein and activates the Gs/adenylyl cyclase pathway to elevate cAMP. Neither the N-terminal ectodomain nor cleavage at the GPCR proteolysis site is required for G protein signaling. Gs coupling was verified by Gαs siRNA knockdown, Gαs overexpression, chimeric Gq(s4) co-expression routing activity to PLC/IP pathway, and a transmembrane-domain missense mutation that abolished receptor activity without altering cell surface expression. |
siRNA knockdown of Gαs, overexpression of Gαs, chimeric G protein co-expression, missense mutagenesis, cAMP assay |
The Journal of biological chemistry |
High |
22025619
|
| 2014 |
A short peptide sequence within the ectodomain of GPR133 (termed the Stachel sequence) functions as a tethered agonist; upon structural changes in the ectodomain, this intramolecular agonist is exposed to the seven-transmembrane domain to trigger G protein activation. The Stachel sequence shows high receptor specificity. |
Peptide agonist assay, zebrafish Stachel-mutant genetic rescue, exogenous peptide application in hypomorphic gpr126 mutants |
Cell reports |
High |
25533341
|
| 2016 |
GPR133 knockdown reduces CD133+ glioblastoma stem cell prevalence, tumor cell proliferation, and tumorsphere formation in vitro, and markedly reduces tumor xenograft growth in vivo; the GPR133 knockdown phenotype is rescued by forskolin, indicating signaling is mediated through cAMP. GPR133 mRNA is transcriptionally upregulated by hypoxia in a HIF-1α-dependent manner. |
shRNA knockdown, tumorsphere assay, mouse xenograft implantation, forskolin rescue, HIF-1α-dependent transcription analysis |
Oncogenesis |
High |
27775701
|
| 2016 |
Functional characterization of naturally occurring ADGRD1 missense variants identified several loss-of-function nsSNPs (A448D, Q600stop, C632fs, A761E, N795K) and one gain-of-function nsSNP (F383S) that significantly increased basal receptor activity. |
Site-directed mutagenesis, cAMP functional assay |
BMC genomics |
Medium |
27516204
|
| 2021 |
GPR133 undergoes autoproteolytic cleavage shortly after protein synthesis; the N-terminal fragment (NTF) and C-terminal fragment (CTF) remain noncovalently associated until the receptor is trafficked to the plasma membrane, where NTF-CTF dissociation occurs. Cleavage-competent WT GPR133 generates significantly more cAMP than the uncleavable H543R mutant. A PAR1-CTF/GPR133-NTF proxy system confirmed that thrombin-induced NTF shedding increases intracellular cAMP, supporting a model where NTF dissociation at the plasma membrane promotes receptor activation. |
Subcellular fractionation, co-immunoprecipitation, uncleavable point mutant (H543R), PAR1 chimeric proxy system with thrombin cleavage, cAMP assay |
The Journal of biological chemistry |
High |
34022221
|
| 2021 |
Adgrd1 is expressed on oviductal epithelium; female mice lacking Adgrd1 are sterile due to failure to relieve the ampullary-isthmic junction (AIJ) restraining mechanism, causing inappropriate retention of embryos in the oviduct. Post-ovulatory attenuation of tubal fluid flow is dysregulated in Adgrd1-deficient mice. The extracellular protein Plxdc2, displayed on cumulus cells, was identified as an activating ligand for Adgrd1 by a large-scale extracellular protein interaction screen. |
Constitutive knockout mice, oviductal fluid flow measurement, large-scale extracellular protein interaction screen (AVEXIS), embryo transit imaging |
Nature communications |
High |
33623007
|
| 2022 |
Cryo-EM structures of ADGRD1 (and ADGRF1) in complex with Gs protein revealed that the stalk region preceding the first transmembrane helix acts as the tethered agonist by forming extensive interactions with the transmembrane domain; an autoproteolysis-deficient ADGRF1 structure showed a cleavage-independent manner of receptor activation. A conserved cascade of inter-helix interaction cores mediates stalk-induced activation. |
Cryo-EM structure determination, mutagenesis, functional signaling assays |
Nature |
High |
35418679
|
| 2022 |
Antibodies targeting the N-terminus of GPR133 increase cAMP in a concentration-dependent manner. This effect requires autoproteolytic cleavage: cells expressing the cleavage-deficient H543R mutant did not respond to antibody stimulation. Antibody treatment promotes release of the autoproteolytically cleaved NTF, supporting the model that NTF dissociation promotes receptor activation. |
Antibody treatment of HEK293T cells and patient-derived GBM cells, cAMP assay, cleavage-deficient mutant (H543R), immunoprecipitation of NTF from conditioned medium |
The Journal of biological chemistry |
Medium |
35447113
|
| 2023 |
PTK7 is an extracellular binding partner of GPR133 in glioblastoma, identified by affinity proteomics. PTK7 binds the autoproteolytically generated NTF of GPR133 and its expression in trans increases GPR133 signaling. This allosteric effect requires GPR133 intramolecular cleavage and PTK7 anchoring in the plasma membrane. PTK7's allosteric action is additive with but topographically distinct from orthosteric Stachel peptide activation. |
Affinity proteomics, co-immunoprecipitation, cAMP assay, cleavage-deficient mutant, PTK7 transmembrane-anchoring requirement test, shRNA knockdown in GBM |
Cell reports |
High |
37354459
|
| 2024 |
ESYT1, a Ca2+-dependent mediator of ER-plasma membrane bridge formation, is an intracellular interactor of GPR133 identified by proximity biotinylation proteomics. ESYT1 knockdown/knockout increases GPR133 signaling; overexpression suppresses it without altering plasma membrane GPR133 levels. The interaction requires the Ca2+-sensing C2C domain of ESYT1. Thapsigargin-mediated cytosolic Ca2+ elevation promotes ESYT1-GPR133 dissociation, relieving the signaling-suppressive effect. |
Proximity biotinylation proteomics (BioID), co-immunoprecipitation, ESYT1 KD/KO and overexpression, domain mutagenesis (C2C), thapsigargin treatment, cAMP assay, GBM tumor growth assay |
Cell reports |
High |
38758649
|
| 2025 |
Constitutive and osteoblast-specific knockouts of Gpr133/Adgrd1 in mice cause reduced cortical bone mass and trabecularization characteristic of osteoporosis, due to impaired osteoblast function and increased osteoclast activity. GPR133/ADGRD1 regulates osteoblast differentiation through a combined mechanism involving PTK7 interaction and mechanical forces (demonstrated by stretch assays in vitro and mechanical loading in vivo). Downstream signaling proceeds via cAMP-dependent activation of the β-catenin pathway. Pharmacological activation with agonist AP-970/43482503 (AP503) enhances osteoblast function and alleviates osteoporosis in ovariectomized mice. |
Constitutive and osteoblast-specific knockout mice, in vitro stretch assay, in vivo mechanical loading, cAMP assay, β-catenin pathway analysis, pharmacological agonist treatment, ovariectomy osteoporosis model |
Signal transduction and targeted therapy |
High |
40583059
|
| 2025 |
The small molecule GL64, identified as a selective ADGRD1 agonist, activates ADGRD1 by mimicking the Stachel sequence. ADGRD1 negatively regulates osteoclastogenesis via the cAMP-PKA-NFATC1 signaling pathway, and GL64 administration prevents bone loss in an ovariectomy mouse model. |
Small-molecule agonist identification, cAMP assay, PKA and NFATC1 pathway analysis, osteoclast differentiation assay, ovariectomy mouse model |
Science advances |
High |
40644539
|
| 2025 |
GPR133 exhibits constitutive self-activation via its Stachel sequence and can activate downstream G13 signaling in addition to Gs. A cryo-EM structure of the GPR133-GAIN-miniGα13 complex was resolved at 3.51 Å, revealing both conserved and distinct features compared to the previously resolved GPR133-CTF-Gs complex. |
Cryo-EM structure determination (3.51 Å), in vitro reconstitution of GPR133-GAIN-miniGα13 complex, G13 signaling assay |
Biochemical and biophysical research communications |
Medium |
40570642
|
| 2024 |
GPR133 upregulation in decidual macrophages (caused by promoter hypomethylation) impairs phagocytic function; GPR133 knockdown in THP-1 macrophages significantly improves phagocytic function. |
5-Aza-dC demethylation, siRNA/shRNA knockdown, phagocytosis assay in decidual macrophages and THP-1 cells |
Epigenetics |
Medium |
38564758
|
| 2025 |
ADGRD1 promotes differentiation of adipose progenitor cells (APCs) in vitro and in vivo. In an obese mouse model (high-fat diet), gain-of-function and loss-of-function studies validated that ADGRD1 promotes adipogenesis and improves metabolic homeostasis. Transcription factors MEF2D and TCF12 were identified as regulators of ADGRD1 expression. |
Single-nucleus sequencing trajectory inference, primary APC differentiation assay, gain- and loss-of-function in HFD mouse model, ChIP-seq and RNA-seq analysis |
Science China. Life sciences |
Medium |
39821834
|