Relationship Between the Dimerization of Thyroglobulin and its Ability to Form Triiodothyronine

Authors

Cintia E. Citterio, Chapman UniversityFollow
Yoshiaki Morishita, University of Michigan, Ann Arbor
Nada Dakka, University of Michigan, Ann Arbor
Balaji Veluswamy, University of Michigan, Ann Arbor
Peter Arvan, University of Michigan, Ann Arbor

Document Type

Article

Publication Date

2-12-2018

Abstract

Thyroglobulin (TG) is the most abundant thyroid gland protein, a dimeric iodoglycoprotein (660 kDa). TG serves as the protein precursor in the synthesis of thyroid hormones tetraiodothyronine (T₄) and triiodothyronine (T₃). The primary site for T₃ synthesis in TG involves an iodotyrosine acceptor at the antepenultimate Tyr residue (at the extreme carboxyl terminus of the protein). The carboxyl-terminal region of TG comprises a cholinesterase-like (ChEL) domain followed by a short unique tail sequence. Despite many studies, the monoiodotyrosine donor residue needed for the coupling reaction to create T₃ at this evolutionarily conserved site remains unidentified. In this report, we have utilized a novel, convenient immunoblotting assay to detect T₃ formation after protein iodination in vitro, enabling the study of T₃ formation in recombinant TG secreted from thyrocytes or heterologous cells. With this assay, we confirm the antepenultimate residue of TG as a major T₃-forming site, but also demonstrate that the side chain of this residue intimately interacts with the same residue in the apposed monomer of the TG dimer. T₃ formation in TG, or the isolated carboxyl-terminal region, is inhibited by mutation of this antepenultimate residue, but we describe the first substitution mutation that actually increases T₃ hormonogenesis by engineering a novel cysteine, 10 residues upstream of the antepenultimate residue, allowing for covalent association of the unique tail sequences, and that helps to bring residues Tyr²⁷⁴⁴ from apposed monomers into closer proximity.

Comments

This article was originally published in Journal of Biological Chemistry, volume 293, issue 13, in 2018. https://doi.org/10.1074/jbc.RA118.001786

Recommended Citation

C. E. Citterio, Y. Morishita, N. Dakka, B. Veluswamy, and P. Arvan (2018). Relationship Between the Dimerization of Thyroglobulin and its Ability to Form Triiodothyronine. J Biol Chem. 293 (13) 4860-4869. https://doi.org/10.1074/jbc.RA118.001786

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