A receptor binding site identified in the region 81-95 of the beta-subunit of human luteinizing hormone (LH) and chorionic gonadotropin (hCG).
Two series of overlapping peptides comprising the entire sequences of the beta-subunits of human lutropin (LH) and choriogonadotropin (hCG) were prepared by a comprehensive synthetic strategy in order to identify all linear regions of the subunit that may participate in binding of the hormone to its receptor. Each series of peptides (15 residues in length) spanned the entire amino acid sequences of the two beta-subunits. The peptides were tested for their ability to inhibit the binding of 125I-labeled hCG or LH to rat ovarian membranes and for their ability to inhibit hCG-stimulated progesterone production in a Leydig cell bioassay. The most potent inhibitor of LH/hCG binding was a peptide containing the sequence beta 81-95, a receptor binding site of the LH/hCG beta subunit not previously described. The concentration at which LH/hCG binding was inhibited at 50% (IC50) was 20 microM and 30 microM for hCGbeta 81-95 and LH beta 81-95, respectively. These peptides also inhibited the stimulation of progesterone production by hCG in Leydig cell bioassays. In order to determine important residues that inhibit binding within this region, a third set of peptides was synthesized in which each residue of hCG beta 81-95 was sequentially replaced with the residue L-alanine. Five residues (Leu-86, Cys-88, Cys-90, Arg-94, and Arg-95) were critical for maximal inhibition of hCG binding by CG beta 81-95. In addition to site beta 81-95, other sites that inhibited hCG/LH binding but with significantly lower potencies included hCG beta 1-15, LH beta 41-55, and LH beta 91-105.(ABSTRACT TRUNCATED AT 250 WORDS)
Morbeck
D E
DE
Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN 55905.
Roche
P C
PC
Keutmann
H T
HT
McCormick
D J
DJ
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D006063:Chorionic Gonadotropin / Q000737:chemistry*; D005260:Female; D007984:Leydig Cell Tumor / Q000378:metabolism / Q000473:pathology; D007986:Luteinizing Hormone / Q000737:chemistry*; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000378:metabolism; D010053:Ovary / Q000378:metabolism; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D011869:Radioligand Assay; D051381:Rats; D011974:Receptors, LH / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D006063:Chorionic Gonadotropin; D009363:Neoplasm Proteins; D010446:Peptide Fragments; D011974:Receptors, LH; D007986:Luteinizing Hormone