title: Binding sites for growth hormone in rabbit placental cytosol.
abstract: A soluble GH-binding protein, which cross-reacted with a monoclonal antibody (Mab) to the rabbit liver membrane GH receptor, has been identified in cytosol preparations from both fetal and maternal portions of rabbit placenta. Structural studies using gel filtration chromatography and chemical covalent cross-linking techniques have shown that the GH-binding protein in fetal/maternal placental cytosol has a native mol wt of 104,000 and a denatured subunit of about 57,000 mol wt (with or without dithiothreitol). Very low levels of GH-specific [125I]human (h) GH binding were observed in membrane preparations from the corresponding placentae, even after desaturation of any endogenously bound hormone by 5 M MgCl2. No PRL-specific binding was observed in placental membranes or cytosols. Scatchard analysis of [125I]hGH binding to fetal and maternal placental cytosol revealed linear plots with Ka values of 6.1 +/- 1.1 nM-1 (fetal) and 5.31 +/- 0.63 nM-1 (maternal; mean +/- SEM; n = 5). The binding capacity of maternal placental cytosol, when expressed as femtomoles per mg protein (170 +/- 10.10) or femtomoles per g tissue (3245 +/- 123), was about 3-fold higher than that for fetal placental cytosol. Northern blot analysis of fetal and maternal placental mRNA probed with a GH receptor oligonucleotide probe revealed hybridization to a 4.4 to 4.7-kilobase and a 2.2-kilobase species in fetal placenta only. The level of GH-specific binding observed in fetal and maternal placental cytosol did not correlate directly with the level of mRNA expression. A GH-binding protein has also been shown to be present in fetal rabbit serum and is known to be structurally and immunologically related to the rabbit placental and liver cytosolic GH-binding proteins. Scatchard analysis of [125I]hGH binding to fetal serum GH-binding protein revealed a single class of high affinity sites with a Ka of 4.64 +/- 1.29 nM-1 and a capacity of 338 +/- 167 fmol/ml serum (mean +/- SEM; n = 4). Given the relative binding capacities and the demonstration of GH receptor mRNA in fetal placental cytosol, it is highly unlikely that contamination of fetal placental cytosol by fetal serum accounts for all of the placental binding capacity observed. However, no such definitive conclusion regarding contamination by maternal rabbit serum of maternal placental cytosol can be made. The presence of GH-binding proteins in placental cytosol has not been described previously, and these observations suggest that GH may have a role in placental metabolism.
authors.affiliation: Medical Research Centre, Prince Henry's Hospital Campus, Monash Medical Centre, Melbourne, Victoria, Australia.
authors.forename: S I
authors.initials: SI
authors.lastname: Ymer
authors.forename: J L
authors.initials: JL
authors.lastname: Stevenson
authors.forename: A C
authors.initials: AC
authors.lastname: Herington
mesh_terms: D000818:Animals; D001345:Autoradiography; D001665:Binding Sites; D015152:Blotting, Northern; D003432:Cross-Linking Reagents / Q000494:pharmacology; D003600:Cytosol / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D013006:Growth Hormone / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D010920:Placenta / Q000378:metabolism* / Q000648:ultrastructure; D011247:Pregnancy; D011485:Protein Binding; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D011986:Receptors, Somatotropin / Q000235:genetics / Q000378:metabolism
chemical_list: D003432:Cross-Linking Reagents; D012333:RNA, Messenger; D011986:Receptors, Somatotropin; D013006:Growth Hormone