HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via ISI Web of Science (3) Citing Articles via Google Scholar Google Scholar Articles by Neumann, S. Articles by Paschke, R. Search for Related Content PubMed PubMed Citation Articles by Neumann, S. Articles by Paschke, R.

Interactions between the extracellular domain and the extracellular loops as well as the 6th transmembrane domain are necessary for TSH receptor activation

III Medical Department, University of Leipzig, Philipp-Rosenthal-Str. 27, 04103 Leipzig, Germany

(Correspondence should be addressed to R Paschke; Email: pasr{at}medizin.uni-leipzig.de)

Objective: The molecular mechanisms of TSH receptor (TSHR) activation and intramolecular signal transduction are largely unknown. Deletion of the extracellular domain (ECD) of the TSHR results in increased constitutive activity, which suggests a self-inhibitory interaction between the ECD and the extracellular loops (ECLs) or the transmembrane domains (TMDs). To investigate these potential interactions and to pursue the idea that mutations in the ECD affect the constitutive activity of mutants in the ECLs or TMDs we generated double mutants between position 281 in the ECD and mutants in all three ECLs as well as the 6th TMD.

Design: We combined mutation S281D, characterized by an impaired TSH-stimulated cAMP response, with the constitutively activating in vivo mutations I486F (1st ECL), I568T (2nd ECL), V656F (3rd ECL) and D633F (6th TMD). Further, we constructed double mutants containing the constitutively activating mutation S281N and one of the inactivating mutations D474E, T477I (1st ECL) and D633K (6th TMD).

Results: The cAMP level of the double mutants with S281N and the inactive mutants in the 1st ECL was decreased below the level of the inactive single mutants, demonstrating that a constitutively activating mutation in the ECD cannot bypass disruption of signal transduction in the serpentine domain. In double mutants with S281D, basal and TSH-induced cAMP and inositol phosphate production of constitutively active mutants was reduced to the level of S281D.

Conclusion: The dominance of S281D and the dependence of constitutively activating mutations in the ECLs on the functionally intact ECD strongly suggest that interactions between these receptor domains are required for TSHR activation and intramolecular signal transduction.

This article has been cited by other articles:

				G. Kleinau, H. Jaeschke, S. Mueller, B. M. Raaka, S. Neumann, R. Paschke, and G. Krause Evidence for cooperative signal triggering at the extracellular loops of the TSH receptor FASEB J, August 1, 2008; 22(8): 2798 - 2808. [Abstract] [Full Text] [PDF]

				S. Mueller, G. Kleinau, H. Jaeschke, S. Neumann, G. Krause, and R. Paschke Significance of Ectodomain Cysteine Boxes 2 and 3 for the Activation Mechanism of the Thyroid-stimulating Hormone Receptor J. Biol. Chem., October 20, 2006; 281(42): 31638 - 31646. [Abstract] [Full Text] [PDF]

				M. Claus, H. Jaeschke, G. Kleinau, S. Neumann, G. Krause, and R. Paschke A Hydrophobic Cluster in the Center of the Third Extracellular Loop Is Important for Thyrotropin Receptor Signaling Endocrinology, December 1, 2005; 146(12): 5197 - 5203. [Abstract] [Full Text] [PDF]