Probing conformational and functional states of human hepatocyte growth factor by a panel of monoclonal antibodies.
HGF-Met signaling contributes to various biological events by controlling cell migration. Since the abnormal activation of Met receptor causes cancer progression, inhibitors such as neutralizing antibodies are regarded as promising therapeutics. HGF is secreted as a single-chain (sc) precursor and is processed by extracellular proteases to generate disulfide-bonded two-chain (tc) HGF. Although this proteolytic processing of HGF is necessary for its biological activity, exactly how the proteolysis leads to the conversion of HGF to the active form is still unclar due to the lack of structural information. In order to gain insights about this point, we generated 6 antibodies against HGF. All antibodies recognized different epitopes on the native HGF protein and showed distinct effects when tested in a cell-based HGF-Met signaling assay. They included one antibody (t1E4) that strongly blocks Met activation by tcHGF, as well as one antibody (t8E4) exclusively recognizing the active tcHGF but not inactive scHGF. Thus, a panel of anti-HGF antibodies suitable for probing the structural mechanism of HGF activation were obtained.
Laboratory of Protein Synthesis and Expression, Institute for Protein Research, Osaka University, Osaka, 565-0871, Japan.
Masataka
M
Umitsu
Division of Tumor Dynamics and Regulation, Cancer Research Institute, Kanazawa University, Ishikawa, 920-1192, Japan.
Katsuya
K
Sakai
Department of Regional Innovation, Tohoku University Graduate School of Medicine, Miyagi, 980-8575, Japan.
Satoshi
S
Ogasawara
Department of Regional Innovation, Tohoku University Graduate School of Medicine, Miyagi, 980-8575, Japan.
Mika K
MK
Kaneko
Laboratory of Protein Synthesis and Expression, Institute for Protein Research, Osaka University, Osaka, 565-0871, Japan.
Ryoko
R
Asaki
Laboratory of Protein Synthesis and Expression, Institute for Protein Research, Osaka University, Osaka, 565-0871, Japan.
Keiko
K
Tamura-Kawakami
Department of Regional Innovation, Tohoku University Graduate School of Medicine, Miyagi, 980-8575, Japan.
Yukinari
Y
Kato
Division of Tumor Dynamics and Regulation, Cancer Research Institute, Kanazawa University, Ishikawa, 920-1192, Japan.
Kunio
K
Matsumoto
Laboratory of Protein Synthesis and Expression, Institute for Protein Research, Osaka University, Osaka, 565-0871, Japan.
Junichi
J
Takagi
D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000737:chemistry* / Q000276:immunology / Q000494:pharmacology; D004285:Dogs; D000939:Epitopes / Q000737:chemistry* / Q000276:immunology; D017228:Hepatocyte Growth Factor / Q000737:chemistry* / Q000276:immunology; D006801:Humans; D061985:Madin Darby Canine Kidney Cells; D051379:Mice; D011487:Protein Conformation; D019859:Proto-Oncogene Proteins c-met / Q000276:immunology; D015398:Signal Transduction / Q000187:drug effects / Q000276:immunology
D058846:Antibodies, Monoclonal, Murine-Derived; D000939:Epitopes; D017228:Hepatocyte Growth Factor; C491743:MET protein, human; D019859:Proto-Oncogene Proteins c-met