The human thyrotropin receptor is predominantly internalized by beta-arrestin 2.
The beta-arrestin-dependent endocytosis of the beta2-adrenergic receptor (beta2AR) has been demonstrated by confocal fluorescence microscopy. Furthermore, a constitutively activated beta2AR is also constitutively desensitized and down-regulated. To clarify the function of beta-arrestin 1 or 2 for TSH receptor (TSHR) desensitization and examine whether constitutively activated TSHR mutants are internalized in a different way, we investigated the TSHR trafficking in association with beta-arrestins in cotransfection experiments in HEK 293 cells using confocal laser-scanning microscopy. We found that both beta-arrestins are able to internalize the TSHR in HEK 293 cells. However, whereas the beta-arrestin 1-mediated TSHR internalization reached its maximum 20 min after TSH stimulation, the beta-arrestin 2-mediated TSHR internalization already reached its maximum 5 min after TSH stimulation. Furthermore, an increased basal desensitization and internalization of constitutively activated TSHR mutants N670S, S505N, and F631L cotransfected with beta-arrestin 2 could not be found. After TSH stimulation the constitutively activated mutants showed the same time course for internalization as the wild-type-TSHR. In summary, contrary to data obtained for the beta2AR, the constitutive activation of the TSHR does not influence the desensitization and time course for internalization of the receptor, and in agreement with findings for the FSH and LH receptors, these results characterize the TSH receptor as a member of the class A of G protein-coupled receptors, which have a higher affinity to beta-arrestin 2 than beta-arrestin 1 and do not colocalize with beta-arrestins in endosomes.
Frenzel
Romy
R
III Medical Department, University of Leipzig, 04103 Leipzig, Germany.
Voigt
Carsten
C
Paschke
Ralf
R
D019390:Arrestins / Q000032:analysis / Q000502:physiology*; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000096:biosynthesis; D011992:Endosomes / Q000378:metabolism; D006801:Humans; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D011989:Receptors, Thyrotropin / Q000032:analysis / Q000378:metabolism*; D015398:Signal Transduction; D013972:Thyrotropin / Q000494:pharmacology; D000071559:beta-Arrestin 1; D000071560:beta-Arrestin 2; D000071557:beta-Arrestins
C000605253:ARRB1 protein, human; C000605254:ARRB2 protein, human; D019390:Arrestins; D043562:Receptors, G-Protein-Coupled; D011989:Receptors, Thyrotropin; D000071559:beta-Arrestin 1; D000071560:beta-Arrestin 2; D000071557:beta-Arrestins; D013972:Thyrotropin; D000242:Cyclic AMP