Hydrodynamic properties of the gonadotropin receptor from a murine Leydig tumor cell line are altered by desensitization.
The murine Leydig tumor cell line 1 (MLTC-1) contains gonadotropin receptors (GR) that are coupled to adenylate cyclase through the stimulatory guanine nucleotide binding protein (Gs). The binding of human choriogonadotropin (hCG) causes MLTC-1 cells to accumulate cAMP. With time, the ability of MLTC-1 cells to respond to hCG is attenuated by a process called desensitization. The hydrodynamic properties of GR from control and desensitized MLTC-1 cells were studied. Sucrose density gradient sedimentation in H2O and D2O and gel filtration chromatography were used to estimate the Stokes radius (a), partial specific volume (vc), sedimentation coefficient (S20,w), and molecular weight (Mr) of the detergent-solubilized hormone-receptor complex (hCG-GR). [125I]hCG was bound to MLTC-1 cells under conditions that allow (37 degrees C) or prevent (0 degree C) desensitization, and hCG-GR was solubilized in Triton X-100. In the absence of desensitization, control hCG-GR had a Mr of 213,000 (a = 6.2; vc = 0.76; S20,w = 7.3), whereas desensitized hCG-GR had a Mr of 158,000 (a = 6.1; Vc = 0.71; S20,w = 6.6). Deglycosylated hCG (DG-hCG) is an antagonist that binds to GR with high affinity but fails to stimulate adenylate cyclase or cause desensitization. [125I]DG-hCG was bound to MLTC-1 cells and DG-hCG-GR solubilized in Triton X-100. The hydrodynamic properties of DG-hCG-GR (Mr 213,000; a = 5.8; Vc = 0.77; S20;w = 7.6) were the same as that for control hCG-GR. There was no evidence for the association of adenylate cyclase or Gs with GR in Triton X-100 solubilized preparations.(ABSTRACT TRUNCATED AT 250 WORDS)
Rebois
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Membrane Biochemistry Section, National Institute of Neurological and Communicative Disorders and Stroke, Bethesda, Maryland 20892.
Bradley
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RM
Titlow
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CC
D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D006063:Chorionic Gonadotropin / Q000378:metabolism; D002850:Chromatography, Gel; D004789:Enzyme Activation; D007700:Kinetics; D007984:Leydig Cell Tumor / Q000378:metabolism*; D051379:Mice; D011487:Protein Conformation; D011967:Receptors, Gonadotropin / Q000302:isolation & purification / Q000378:metabolism*
D006063:Chorionic Gonadotropin; D011967:Receptors, Gonadotropin; D000262:Adenylyl Cyclases