Purification and characterization of the bovine pituitary luteinizing hormone releasing hormone M(r) 60,000 binding protein.
Luteinizing hormone releasing hormone (LHRH) regulates the release of luteinizing hormone and follicle stimulating hormone from the pituitary. This process takes place through interaction with high affinity membrane receptors. In addition LHRH inhibits the growth of several cancer cell lines through the interaction with M(r) 60,000 LHRH receptors. Here we describe the purification to homogeneity of the M(r) 60,000 bovine pituitary LHRH binding protein in amounts allowing N-terminal sequencing and peptide mapping. The procedure describes solubilization of luteinizing hormone releasing hormone receptors from homogenized bovine pituitaries in an active form by using the detergent Triton X-114. The receptors were retained in the Triton X-114 phase during temperature-dependent phase separation. Preparative phase separations were performed directly on solubilized bovine pituitary extracts. SDS-PAGE of the purified LHRH receptor after LHRH-immobilized affinity chromatography showed the presence of a single band with M(r) 60,000. Partial sequencing of this band after trypsin digestion of gel pieces revealed unknown sequences with a possible homology to other receptors including some G-protein coupled receptors.
Christiansen
J
J
Institute of Biochemical Genetics, Copenhagen, Denmark.
Houen
G
G
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010449:Peptide Mapping; D010902:Pituitary Gland / Q000737:chemistry*; D011966:Receptors, LHRH / Q000737:chemistry / Q000302:isolation & purification*
D011966:Receptors, LHRH