This Article Full Text Full Text (PDF) Purchase Article View Shopping Cart Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Curcio-Morelli, C. Articles by Bianco, A. C. Search for Related Content PubMed PubMed Citation Articles by Curcio-Morelli, C. Articles by Bianco, A. C.

In Vivo Dimerization of Types 1, 2, and 3 Iodothyronine Selenodeiodinases

Thyroid Section, Division of Endocrinology (C.C.-M., A.M.Z., B.W.K., S.H., J.W.H., P.R.L., A.C.B.), Diabetes and Hypertension, Brigham and Women’s Hospital and Harvard Medical School, Boston, Massachusetts 02115; and Institute of Experimental Medicine (B.G.), Department of Neurobiology, Budapest and University of Pécs, Faculty of Sciences, Institute of Biology, Budapest H-1083, Hungary

Address all correspondence and requests for reprints to: Antonio C. Bianco, M.D., Ph.D., Brigham and Women’s Hospital, 77 Avenue Louis Pasteur, Harvard Institutes of Medicine Building, Room 566, Boston, Massachusetts 02115. E-mail: abianco{at}rics.bwh.harvard.edu.

The goal of the present investigation was to test the hypothesis that types 1, 2, and 3 iodothyronine selenodeiodinases (D1, D2, and D3) can form homodimers. The strategy included transient coexpression of wild-type (wt) deiodinases (target), and FLAG-tagged alanine or cysteine mutants (bait) in human embryonic kidney epithelial cells. SDS-PAGE of the immunoprecipitation pellet of ⁷⁵Se-labeled cell lysates using anti-FLAG antibody revealed bands of the correct sizes for the respective wt enzymes, which corresponded to approximately 2–5% of the total deiodinase protein in the cell lysate. Western blot analysis with anti-FLAG antibody of lysates of cells transiently expressing individual FLAG-tagged-cysteine deiodinases revealed specific monomeric bands for each deiodinase and additional minor bands of relative molecular mass (M_r) of 55,000 for D1, M_r 62,000 for D2, and M_r 65,000 for D3, which were eliminated by 100 mM dithiothreitol at 100 C. Anti-FLAG antibody immunodepleted 10% of D1 and 38% of D2 activity from lysates of cells coexpressing inactive FLAG-tagged Ala mutants and the respective wt enzymes (D1 or D2) but failed to immunodeplete wtD3 activity. D1 or D2 activities were present in these respective pellets. We conclude 1) that overexpressed selenodeiodinases can homodimerize probably through disulfide bridges; and 2) at least for D1 and D2, monomeric forms are catalytically active, demonstrating that only one wt monomer partner is required for catalytic activity of these two deiodinases.

This article has been cited by other articles:

				B. Gereben, A. M. Zavacki, S. Ribich, B. W. Kim, S. A. Huang, W. S. Simonides, A. Zeold, and A. C. Bianco Cellular and Molecular Basis of Deiodinase-Regulated Thyroid Hormone Signaling Endocr. Rev., December 1, 2008; 29(7): 898 - 938. [Abstract] [Full Text] [PDF]

				G. D. V. Sagar, B. Gereben, I. Callebaut, J.-P. Mornon, A. Zeold, C. Curcio-Morelli, J. W. Harney, C. Luongo, M. A. Mulcahey, P. R. Larsen, et al. The Thyroid Hormone-Inactivating Deiodinase Functions as a Homodimer Mol. Endocrinol., June 1, 2008; 22(6): 1382 - 1393. [Abstract] [Full Text] [PDF]

				J. P Brouwer, B. C Appelhof, R. P Peeters, W. J G Hoogendijk, J. Huyser, A. H Schene, J. G P Tijssen, R. Van Dyck, T. J Visser, W. M Wiersinga, et al. Thyrotropin, but not a polymorphism in type II deiodinase, predicts response to paroxetine in major depression. Eur. J. Endocrinol., June 1, 2006; 154(6): 819 - 825. [Abstract] [Full Text] [PDF]

				G. I. C. Simpson, D. M. Leonard, and J. L. Leonard Identification of the Key Residues Responsible for the Assembly of Selenodeiodinases J. Biol. Chem., May 26, 2006; 281(21): 14615 - 14621. [Abstract] [Full Text] [PDF]

				J. L. Leonard, G. Simpson, and D. M. Leonard Characterization of the Protein Dimerization Domain Responsible for Assembly of Functional Selenodeiodinases J. Biol. Chem., March 25, 2005; 280(12): 11093 - 11100. [Abstract] [Full Text] [PDF]

				C. H. A. Gouveia, M. A. Christoffolete, C. R. Zaitune, J. M. Dora, J. W. Harney, A. L. Maia, and A. C. Bianco Type 2 Iodothyronine Selenodeiodinase Is Expressed throughout the Mouse Skeleton and in the MC3T3-E1 Mouse Osteoblastic Cell Line during Differentiation Endocrinology, January 1, 2005; 146(1): 195 - 200. [Abstract] [Full Text] [PDF]

				G. G. J. M. Kuiper, F. Wassen, W. Klootwijk, H. van Toor, E. Kaptein, and T. J. Visser Molecular Basis for the Substrate Selectivity of Cat Type I Iodothyronine Deiodinase Endocrinology, December 1, 2003; 144(12): 5411 - 5421. [Abstract] [Full Text] [PDF]