Document Type
Article
Publication Date
9-17-2023
Abstract
Thyroglobulin must pass endoplasmic reticulum (ER) quality control to become secreted for thyroid hormone synthesis. Defective thyroglobulin, blocked in trafficking, can cause hypothyroidism. Thyroglobulin is a large protein (~2750 residues) spanning regions I–II–III plus a C-terminal cholinesterase-like domain. The cholinesterase-like domain functions as an intramolecular chaperone for regions I–II–III, but the folding pathway leading to successful thyroglobulin trafficking remains largely unknown. Here, informed by the recent three-dimensional structure of thyroglobulin as determined by cryo-electron microscopy, we have bioengineered three novel classes of mutants yielding three entirely distinct quality control phenotypes. Specifically, upon expressing recombinant thyroglobulin, we find that first, mutations eliminating a disulfide bond enclosing a 200-amino acid loop in region I have surprisingly little impact on the ability of thyroglobulin to fold to a secretion-competent state. Next, we have identified a mutation on the surface of the cholinesterase-like domain that has no discernible effect on regional folding yet affects contact between distinct regions and thereby triggers impairment in the trafficking of full-length thyroglobulin. Finally, we have probed a conserved disulfide in the cholinesterase-like domain that interferes dramatically with local folding, and this defect then impacts on global folding, blocking the entire thyroglobulin in the ER. These data highlight variants with distinct effects on ER quality control, inhibiting domain-specific folding; folding via regional contact; neither; or both.
Recommended Citation
Citterio CE, Kim K, Rajesh B, Pena K, Clarke OB, Arvan P. Structural features of thyroglobulin linked to protein trafficking. Protein Sci. 2023;32(11):e4784. https://doi.org/10.1002/pro.4784
Supplementary information
Copyright
Wiley
Included in
Amino Acids, Peptides, and Proteins Commons, Hormones, Hormone Substitutes, and Hormone Antagonists Commons, Medicinal and Pharmaceutical Chemistry Commons
Comments
This is the accepted version of the following article:
Citterio CE, Kim K, Rajesh B, Pena K, Clarke OB, Arvan P. Structural features of thyroglobulin linked to protein trafficking. Protein Sci. 2023;32(11):e4784. https://doi.org/10.1002/pro.4784
which has been published in final form at https://doi.org/10.1002/pro.4784. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.