| 2000 |
Active HGFA forms a specific complex with membrane-bound HAI-1 (but not HAI-2) on the surface of epithelial cells, requiring HGFA enzymatic activity for binding. HAI-1 acts as both an inhibitor and a cell-surface reservoir of active HGFA. The HGFA·HAI-1 complex is released from the cell surface by phorbol ester or IL-1β treatment via a zinc-metalloproteinase-dependent shedding mechanism, generating 58-kDa HAI-1 fragments with reduced inhibitory potency and restoring HGFA activity in the supernatant. |
Co-immunoprecipitation, CHO cell engineered expression system, cell-surface binding assays, phorbol ester/IL-1β treatment with metalloproteinase inhibitor BB3103 |
The Journal of biological chemistry |
High |
11013244
|
| 2005 |
Crystal structure of the HGFA protease domain at 2.7 Å revealed an unconventional, non-enzymatically competent conformation of active-site residues in the unliganded state. Co-crystal structure with the first Kunitz domain (KD1) of HAI-1B at 2.6 Å showed that inhibitor binding rearranges the 220-loop and 99-loop into a substrate-binding-competent conformation. KD1 occupies subsites S1, S2, and S4 in a substrate-like manner and is solely responsible for the inhibitory specificity of the HAI-1B extracellular region. HGFA, matriptase, hepsin, plasma kallikrein, and trypsin are potently inhibited by KD1. |
X-ray crystallography (2.7 Å apo, 2.6 Å complex), serine protease inhibition panel assay |
Journal of molecular biology |
High |
15713485
|
| 2008 |
Kallikrein-related peptidases KLK4 and KLK5 cleave pro-HGFA at the normal processing site Arg407-Ile408, activating the zymogen. KLK5 required a negatively charged substance (dextran sulfate) for efficient processing whereas KLK4 did not. HGFA activated by these KLKs efficiently converted pro-HGF/SF into active form, leading to cellular scattering and invasion in vitro. Both KLK4 and KLK5 activity on pro-HGFA was strongly inhibited by HAI-1. |
In vitro protease cleavage assay, N-terminal sequencing of cleavage site, cell scattering/invasion assay, inhibitor (HAI-1) functional assay |
The FEBS journal |
High |
18221492
|
| 2010 |
Crystal structures of the HGFA short-form (34 kDa protease domain) reveal two conformational states of substrate-specificity-determining loops (220-loop and 99-loop): enzymatically competent and non-competent. In the KD1-HGFA complex structure, KD1 side chains occupying S1, S2, and S4 subsites are virtually superimposable on the P1, P2, and P4 residues of a pro-HGF-derived substrate mimic (Lys-Gln-Leu-Arg chloromethyl ketone), rationalizing HGFA's narrow substrate specificity for pro-HGF and pro-macrophage-stimulating protein. |
X-ray crystallography, substrate mimic chloromethyl ketone co-crystal comparison |
The FEBS journal |
High |
20402765
|
| 2010 |
Protein C inhibitor (PCI/SERPINA5) forms a complex with active HGFA and inhibits HGFA-catalyzed activation of pro-HGF. In hPCI-transgenic mice, liver regeneration after partial hepatectomy was significantly impaired compared to wild-type, and this impairment was rescued by anti-human PCI antibody. Plasma HGFA-PCI complex levels were elevated in patients after hepatectomy. |
In vitro complex formation assay, hPCI-transgenic mouse partial hepatectomy model, antibody rescue experiment, patient plasma ELISA |
The FEBS journal |
Medium |
20402764
|
| 2010 |
HGFA is a serine protease activated downstream of the blood coagulation cascade (thrombin cleaves pro-HGFA), linking tissue injury to proteolytic activation of pro-HGF. HGFA also activates macrophage-stimulating protein (MSP), the ligand for RON receptor. HGFA-knockout mice show impaired regeneration of severely damaged mucosal epithelium and impaired initial macrophage recruitment in injured tissue in vivo. |
HGFA-knockout mouse model, in vivo tissue injury/regeneration assays |
The FEBS journal |
Medium |
20402763
|
| 2017 |
Systemic active HGFA, released in response to injury, is sufficient to induce transition of skeletal muscle stem cells (MuSCs) and fibro-adipogenic progenitors (FAPs) into GAlert state via proteolytic processing/activation of HGF. Administration of active HGFA to animals accelerated stem cell activation and tissue repair. |
In vivo systemic HGFA administration to mice, stem cell functional activation assays, tissue repair readouts |
Cell reports |
Medium |
28423312
|
| 2012 |
HNF1α directly regulates transcription of the Hgfac gene in pancreatic β-cells. HNF1α knockdown in MIN6 cells decreased Hgfac expression, and Hgfac expression was also reduced in islets of HNF1α(+/-) mice. Reporter gene analysis and chromatin immunoprecipitation (ChIP) confirmed direct HNF1α binding to the Hgfac promoter. |
shRNA knockdown, reporter gene assay, chromatin immunoprecipitation (ChIP), HNF1α heterozygous mouse model |
Biochemical and biophysical research communications |
Medium |
22877752
|
| 2023 |
ChREBP directly regulates HGFAC transcription in liver (identified by ChREBP ChIP-Seq). HGFAC-KO mice exhibited metabolic phenotypes concordant with human loss-of-function variants. In gain- and loss-of-function mouse models, HGFAC enhanced lipid and glucose homeostasis, potentially mediated through activation of hepatic PPARγ activity. |
ChREBP ChIP-Seq, HGFAC knockout mouse, gain-of-function genetic mouse models, metabolic phenotyping, PPARγ activity assay |
JCI insight |
Medium |
36413406
|
| 2023 |
DNMT3A methylates the HGFAC promoter, silencing HGFAC expression in hepatocellular carcinoma. miR-4270 targets the 3'UTR of DNMT3A (confirmed by dual-luciferase and Ago2-RIP assays), reducing DNMT3A-mediated methylation of the HGFAC promoter (confirmed by ChIP and methylation-specific PCR), thereby restoring HGFAC expression. HGFAC overexpression counteracted HCC cell growth promoted by miR-4270 inhibition. |
Dual-luciferase reporter assay, Ago2-RIP, Co-IP, ChIP, methylation-specific PCR, rescue functional assays, xenograft mouse model |
PeerJ |
Medium |
38077422
|
| 2014 |
Substrate-based ketothiazole inhibitors (e.g., Ac-KRLR-kt) inhibit HGFA enzymatic activity at nM Ki values (as low as 12 nM), block HGFA-mediated conversion of native pro-HGF and pro-MSP, and cause dose-dependent decrease of c-MET signaling in MDA-MB-231 breast cancer cells. |
Kinetic fluorogenic enzyme assay, native pro-HGF/pro-MSP cleavage assay, c-MET phosphorylation in cancer cells |
ACS medicinal chemistry letters |
Medium |
25408834
|
| 2001 |
HGFA (factor XII-like serine protease) critically mediates the enhanced activation of HGF/SF observed in colorectal carcinoma tissues compared with normal mucosa, as assessed by the ratio of two-chain (active) to single-chain (latent) HGF. HAI-1 functions both as a cell-surface specific inhibitor of active HGFA and as a reservoir/acceptor on the cell surface, concentrating pericellular HGFA activity. |
Western blot analysis of HGF activation state in tumor vs. normal tissue, cell-surface binding assays, IL-1β-induced shedding assay |
Human cell |
Low |
11436357
|