Characterization and structure of ovarian and testicular LH/hCG receptors.
Ovarian and Leydig cell LH/hCG receptors purified to homogeneity were identified as a single protein of Mr 80,000 and 90,000 respectively. The homogeneity of this protein was confirmed by microsequencing of the first 18 amino acids of the ovarian receptor. The unblocked N-terminal peptide consisted of NH2-R-E-L-S-G-S-R-X-P-E-P-X-D-X-A-P-D-G. These receptors are N-linked sialoglycoproteins which accounts for the size difference between testicular and ovarian receptors and may participate in the interaction with gonadotropin. Crosslinking of pure receptor with hCG with 125I label in either subunit indicated significant interaction of alpha-hCG with the receptor, while beta-hCG seems involved mostly through association and conformational influence on the alpha-subunit. Comparison of Mr derived from SDS with those from FPLC suggested that the native LH receptor are dimers of identical subunits. Autoradiographs of blotted receptors demonstrated that both monomeric and dimeric forms can bind hCG. Receptors from both tissues can be phosphorylated by the catalytic subunit of cAMP-dependent protein kinase and phosphopeptide maps were identical. Receptor occupancy by agonist leads to a conformational change which facilitates its phosphorylation during initial binding and reduces the rate of phosphorylation after more prolonged exposure to gonadotropin. Aggregation or dimerization of the hCG/LH receptors could promote clustering and or crosslinking of receptors in the membrane favouring the initial transduction steps in the action of these hormones.
Dufau
M L
ML
Section on Molecular Endocrinology, National Institute of Child Health and Human Development, Bethesda, Maryland 20892.
Minegishi
T
T
Buczko
E S
ES
Delgado
C J
CJ
Zhang
R
R
D000595:Amino Acid Sequence; D000818:Animals; D001345:Autoradiography; D015153:Blotting, Western; D002850:Chromatography, Gel; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D007985:Leydig Cells / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D010053:Ovary / Q000166:cytology / Q000378:metabolism*; D010766:Phosphorylation; D051381:Rats; D011974:Receptors, LH / Q000302:isolation & purification*; D013737:Testis / Q000378:metabolism*
D003432:Cross-Linking Reagents; D011974:Receptors, LH