Desensitization and internalization of a thyrotrophin receptor lacking the cytoplasmic carboxy-terminal region.
To understand the functional significance of the carboxy-terminal half of the intracellular region of the human TSH receptor (hTSHR), a mutant hTSHR lacking amino acids from the carboxy-terminal to His726 was constructed. Wild type hTSHR cDNA and truncated hTSHR cDNA were subcloned into a eukaryotic expression vector, pRc/CMV, and transfected into Chinese hamster ovary cells to obtain cell lines which stably expressed hTSHRs at high levels. This allowed us to observe highly efficient coupling of hTSHR and adenylyl cyclase as well as desensitization and internalization of hTSHR. Despite the differences in potential phosphorylation sites and internalization signals, dose-dependent stimulation of adenylyl cyclase by TSH, TSH-dependent desensitization and the rate of hTSHR internalization were similar for wild type and truncated hTSHRs. We conclude that the carboxy-terminal half of the intracellular region of hTSHR does not have a major functional role in TSH-dependent signal transduction.
Haraguchi
K
K
Third Department of Internal Medicine, University of Yamanashi Medical School, Japan.
Saito
T
T
Kaneshige
M
M
Endo
T
T
Onaya
T
T
D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D006224:Cricetinae; D000242:Cyclic AMP / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D011485:Protein Binding; D011989:Receptors, Thyrotropin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion; D015398:Signal Transduction; D013972:Thyrotropin / Q000378:metabolism / Q000494:pharmacology
D011989:Receptors, Thyrotropin; D013972:Thyrotropin; D000242:Cyclic AMP