| 2013 |
The KRIT1 FERM domain binds both Rap1 GTPase and the HEG1 cytoplasmic tail simultaneously; crystal structure of the KRIT1-Rap1-HEG1 ternary complex revealed that HEG1 binds in a hydrophobic pocket at the KRIT1 F1-F3 interface with no overlap with the Rap1-binding site (Kd ~1.2 µM), and Rap1 binds on the F1-F2 surface. A KRIT1(K570I) mutant with 8-fold reduced Rap1 affinity confirmed the specific ionic interaction between the F2 lobe and Rap1. |
Crystal structure of ternary complex, surface plasmon resonance / binding affinity measurements, mutagenesis (KRIT1 K570I) |
The Journal of biological chemistry |
High |
23814056
|
| 2021 |
A small-molecule inhibitor (HKi2) competes orthosterically with the HEG1 cytoplasmic tail for the KRIT1 FERM domain binding pocket; crystal structure of HKi2-KRIT1 FERM confirmed it occupies the same pocket as HEG1. Acute inhibition of the HEG1-KRIT1 interaction in human endothelial cells increased KLF4 and KLF2 mRNA and protein levels and triggered PI3K-dependent Akt phosphorylation. |
Crystal structure of inhibitor-KRIT1 complex, in vitro colocalization displacement assay, siRNA knockdown, pharmacological inhibition in endothelial cells, genome-wide RNA-seq, zebrafish in vivo reporter |
FASEB bioAdvances |
High |
33977234
|
| 2018 |
Zebrafish Heg1 stabilizes Krit1 protein levels; Heg1 expression is positively regulated by blood flow; both Heg1 and Krit1 dampen expression of the mechanosensitive gene klf2a, and loss of Krit1 causes increased klf2a and notch1b expression throughout the endocardium, preventing cardiac valve leaflet formation. |
Zebrafish genetic loss-of-function (mutants/morpholinos), in vivo reporter assays for klf2a and notch1b, Western blot for Krit1 protein levels, blood-flow manipulation experiments |
eLife |
High |
29364115
|
| 2013 |
Ccm2l (a novel CCM2 paralog) binds CCM1/KRIT1 directly, and CCM2 overexpression can partially rescue ccm2l morphant cardiovascular defects. Deletion and mutational analyses mapped the regions of CCM1 that mediate its binding to Ccm2l, CCM2, and HEG1, placing ccm2l as a component of the Heg-CCM pathway in cardiovascular development. |
Morpholino knockdown in zebrafish, genetic epistasis (morpholino co-injection), biochemical binding/deletion analysis (pulldown/Co-IP) |
Developmental biology |
Medium |
23328253
|
| 2017 |
HEG1 is a sialylated mucin-like membrane protein (~400 kDa); gene silencing of HEG1 significantly suppressed the survival and proliferation of mesothelioma cells, indicating HEG1 supports mesothelioma cell viability. |
siRNA gene silencing in mesothelioma cell lines, cell viability/proliferation assays, monoclonal antibody characterization (SKM9-2) |
Scientific reports |
Medium |
28361969
|
| 2018 |
The SKM9-2 epitope on HEG1 was identified as the O-glycosylated region 893-SKSPSLVSLPT-903; the SKxPSxVS sequence is essential for antibody recognition. Mass spectrometry and lectin analysis showed the epitope contains two disialylated core 1 O-linked glycan-modified serine residues (Ser893 and Ser900), while Ser897 is not glycosylated. |
Truncated HEG1 binding assays, alanine scanning mutagenesis, mass spectrometry, lectin binding analysis, neuraminidase treatment |
Scientific reports |
High |
30250045
|
| 2023 |
Stable flow induces HEG1 mRNA and protein expression in endothelial cells, promotes HEG1 translocation to the downstream side of cells and release into the media. HEG1 knockdown prevents stable flow-induced KLF2/4 expression by regulating KRIT1 and the MEKK3-MEK5-ERK5-MEF2 pathway. Endothelial-specific HEG1 knockout mice develop exacerbated atherosclerotic plaques. |
siRNA knockdown in human aortic endothelial cells under defined flow conditions, endothelial-specific inducible knockout mice (HEG1iECKO), partial carotid ligation model, Western blot, qPCR, immunostaining |
Circulation |
High |
38099436
|
| 2025 |
HEG1 regulates NO bioavailability via a flow-dependent direct interaction with eNOS (NOS3) in endothelial cells. Endothelial-specific Heg1 knockout mice develop spontaneous hypertension and severe atherosclerosis, both of which are prevented by ACE inhibition. |
Endothelial-specific Heg1 knockout mice, co-immunoprecipitation (HEG1-eNOS interaction), blood pressure monitoring, ACE inhibitor treatment |
bioRxivpreprint |
Medium |
40667131
|
| 2026 |
Endothelial HEG1 facilitates CUL3-mediated proteasomal degradation of PHACTR1. HEG1 deletion leads to increased PHACTR1 levels, its nuclear translocation, and suppression of SP1-mediated eNOS transcription and NO production, resulting in impaired vasodilation and hypertension. |
Endothelial-specific Heg1 deletion mice, proteomics, transcriptomics, ubiquitination assays, pharmacological PHACTR1 inhibition (CCG-1423), vascular function analysis |
European heart journal |
Medium |
40986512
|
| 2022 |
Liver endothelial Heg deficiency reduces expression of Wnt ligands/agonists (Wnt2, Wnt9b, Rspo3) in endothelial cells, limiting Axin2-mediated canonical Wnt signaling and cytochrome P450 expression in hepatocytes, thereby altering liver metabolic zonation. |
Global (Heg-/-) and liver endothelial-specific (Lyve1-Cre;Hegfl/fl) conditional knockout mice, RNA-seq, histology, biochemical assays, 3D vascular network visualization |
Cellular and molecular gastroenterology and hepatology |
High |
35202885
|
| 2026 |
Endothelial Heg1 deletion exacerbates hepatic steatosis under metabolic stress by downregulating BMP signaling, reducing Wnt2, Wnt9b, and Rspo3 expression in endothelial cells, which attenuates hepatocyte Wnt signaling, decreases PPARα and fatty acid oxidation enzyme expression. Restoration of RSPO3 in endothelial cells reversed the steatotic phenotype. |
Two endothelial-specific Heg1 knockout models, three liver disease models (HFD, MCD diet, alcohol), transcriptomics, lipidomics, RSPO3 conditional knock-in rescue, BMP activator treatment |
Cellular and molecular gastroenterology and hepatology |
High |
42119921
|
| 2024 |
Zebrafish Heg1 is dynamically localized at endothelial cell-cell junctions during anastomosis and is required for oscillatory actomyosin contractility along junctions. In heg1 and krit1 mutants, cell-cell interfaces become entangled due to lack of junctional actomyosin contractility, preventing continuous lumen formation; optogenetic RhoA activation restores junction straightening in mutants. |
Live imaging with Heg1 and Myosin reporters in zebrafish, CRISPR/Cas9 mutants, optogenetic RhoA activation, confocal microscopy |
Angiogenesis |
High |
39249713
|
| 2025 |
HEG1 stabilizes AKT1 by reducing its ubiquitination in gastric cancer cells, leading to sustained AKT signaling. The deubiquitinase USP48 directly interacts with HEG1 and stabilizes it by removing K48-linked polyubiquitin chains, preventing its proteasomal degradation. |
Immunoprecipitation, ubiquitination assays (K48-linked polyubiquitin chain analysis), siRNA/overexpression in gastric cancer cells, in vivo tumor xenograft, RNA-seq |
European journal of medical research |
Medium |
41013721
|
| 2025 |
HEG1 protein is decorated with low-sulfated keratan sulfate (KS) O-glycans in malignant pleural mesothelioma; the KS modification is carried on the HEG1 core protein as demonstrated by western blot of HEG1·IgG recombinant fusion proteins secreted from KS-expressing cells, and sensitivity to endo-β-galactosidase and keratanase II but not PNGase F confirmed O-linked glycan attachment. |
Immunohistochemistry with endoglycosidase treatments, reversed-phase ion-pair HPLC disaccharide analysis, western blot of recombinant HEG1·IgG glycoforms |
Pathology international |
Medium |
40525709
|
| 2020 |
HEG1 suppression in malignant mesothelioma cell lines reduces cell proliferation and induces apoptosis; microRNA-23b (miR-23b) acts downstream of HEG1 to support cell proliferation by suppressing apoptosis and autophagy (LC3-II induction). Combined inhibition of miR-23b and HEG1 showed additive antiproliferative effects. |
siRNA knockdown, miRNA inhibitor transfection in mesothelioma cell lines, MTS assay, Annexin V apoptosis assay, western blot (LC3-II) |
Biochemical and biophysical research communications |
Medium |
32284171
|