title: Differential intracellular signalling properties of the growth hormone receptor induced by the activation of an anti-GHR antibody.
abstract: A series of studies have reported that anti-GHR antibody can function as a GHR agonist and may serve as an attractive tool for studying the mechanisms of GHR activation. However, to date, there is relatively little information about intracellular signalling triggered by anti-GHR antibody. Therefore, in this work, we have developed a panel of monoclonal antibodies to GHBP, among which one Mab, termed CG-172, was selected for further characterisation because of its signalling properties. The results from FACS assays, receptor binding and immunoprecipitation assays and western blotting demonstrated that CG-172 specifically binds to GHR expressed on target cells. Subsequently, epitope mapping studies that used receptor binding analysis showed that CG-172 specifically binds subdomain 1 of GHR ECD. We next examined the resulting signal transduction pathways triggered by this antibody in CHO-GHR638 cells and rat hepatocytes. We found that CG-172 can activate JAK2, AKT, ERK1/2 and STAT1/3 but not STAT5. The phosphorylation kinetics of STAT1/3, AKT and ERK1/2 induced by either GH or CG-172 were analysed in dose-response and time course experiments. Our observations demonstrated that an anti-GHR monoclonal antibody (CG-172) can serve as an attractive tool to study the mechanism(s) of GHR-mediated intracellular signalling pathways and may lead to the production of signal-specific molecules that are capable of inducing different biochemical responses.
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin Xincheng Street 2888, Changchun 130118, PR China.
authors.forename: Hainan
authors.initials: H
authors.lastname: Lan
authors.affiliation: State Key Laboratory of Animal Nutrition, China Agricultural University, Beijing 100193, PR China.
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin Xincheng Street 2888, Changchun 130118, PR China.
authors.forename: Zhiling
authors.initials: Z
authors.lastname: Fu
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin Xincheng Street 2888, Changchun 130118, PR China.
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Yang
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin Xincheng Street 2888, Changchun 130118, PR China.
authors.forename: Tiancheng
authors.initials: T
authors.lastname: Wu
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin Xincheng Street 2888, Changchun 130118, PR China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Liu
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin Xincheng Street 2888, Changchun 130118, PR China.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin Xincheng Street 2888, Changchun 130118, PR China.
authors.forename: Huanzhong
authors.initials: H
authors.lastname: Cui
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin Xincheng Street 2888, Changchun 130118, PR China.
authors.forename: Yumeng
authors.initials: Y
authors.lastname: Li
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin Xincheng Street 2888, Changchun 130118, PR China.
authors.forename: Pan
authors.initials: P
authors.lastname: Hong
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin Xincheng Street 2888, Changchun 130118, PR China.
authors.forename: Jingsheng
authors.initials: J
authors.lastname: Liu
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin Xincheng Street 2888, Changchun 130118, PR China. Electronic address: zhengtougao@163.com.
authors.forename: Xin
authors.initials: X
authors.lastname: Zheng
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000494:pharmacology*; D016466:CHO Cells; D049109:Cell Proliferation / Q000187:drug effects; D006224:Cricetinae; D003412:Cricetulus; D018604:Epitope Mapping; D005260:Female; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism; D019382:Human Growth Hormone / Q000494:pharmacology; D006801:Humans; D020935:MAP Kinase Signaling System; D008807:Mice, Inbred BALB C; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D051381:Rats; D011986:Receptors, Somatotropin / Q000819:agonists* / Q000276:immunology / Q000378:metabolism; D050791:STAT Transcription Factors / Q000378:metabolism
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D011986:Receptors, Somatotropin; D050791:STAT Transcription Factors; D019382:Human Growth Hormone; D051057:Proto-Oncogene Proteins c-akt
keywords: Anti-GHR antibody; Growth hormone receptor; Signal transduction pathway