22- and 20 kDa-human growth hormones bind to different sites within certain cellular receptors.
Human growth hormone 20 kDa (20 kDa-hGH) is a natural variant of the main hGH isoform (22 kDa-hGH). Since some 20- and 22 kDa-hGH biological activities are not identical, we decided to map the prolactin (PRLR) and growth hormone receptor (GHR) binding sites for both isoforms. Monoclonal antibody (MAb) R7B4, directed to both receptors, was employed to estimate the relative proximity between 20- and 22 kDa-hGH receptors binding sites. Results indicated that although both hGH isoforms share the same PRLR present in Nb2-cells and rat liver, MAb R7B4 differently affected hormone binding, suggesting that their receptor binding sites would be close in Nb2-cells and separate in rat liver membranes. Since labelled 20 kDa-hGH did not bind significantly to hGHR, we added to the incubating medium an allosteric MAb anti-hGH that improved 20 kDa-hGH affinity for receptors. Under these experimental conditions MAb R7B4 inhibited 20 kDa-hGH binding to human liver but not to placenta, whereas the Ab impaired 22 kDa-hGH binding to both receptors. Data thus suggested that both hGH isoforms share the same hGHR binding site in liver tissue but bind to different overlapped regions in placenta. Consequently, results presented in this paper indicate that PRLR and GHR binding sites for 22- and 20 kDa-hGH should not be always identical, a fact that could explain some of the isoforms different activities.
Longhi
Silvia A
SA
Instituto de Química y Fisicoquímica Biológicas (UBA-CONICET), Facultad de Farmacia y Bioquímica, Buenos Aires, Argentina.
Cortés
María M
MM
Retegui
Lilia A
LA
D000818:Animals; D000911:Antibodies, Monoclonal; D001665:Binding Sites; D002417:Cattle; D005260:Female; D019382:Human Growth Hormone / Q000378:metabolism*; D006801:Humans; D008099:Liver / Q000378:metabolism; D008970:Molecular Weight; D010920:Placenta / Q000378:metabolism; D010928:Placental Lactogen / Q000378:metabolism; D011247:Pregnancy; D011485:Protein Binding; D020033:Protein Isoforms; D051381:Rats; D011981:Receptors, Prolactin / Q000378:metabolism*; D011986:Receptors, Somatotropin / Q000378:metabolism*; D012756:Sheep
D000911:Antibodies, Monoclonal; D020033:Protein Isoforms; D011981:Receptors, Prolactin; D011986:Receptors, Somatotropin; D019382:Human Growth Hormone; D010928:Placental Lactogen