Thyroglobulin (Tg), the prothyroid hormone, is stored in the lumen of the thyroid follicles as soluble dimers and tetramers and insoluble multimers. Soluble Tg is well characterized with regards to structure and role, but insoluble Tg (i-Tg) is not. Here we show that i-Tg, multimerized through formation of disulfide and dityrosine bonds, has a higher iodine content than soluble Tg and no thyroid hormones. Furthermore, the size and the resistance of i-Tg to proteolytic enzymes implied a new mechanism by which thyrocytes may degrade this form of Tg. Using peroxidase and H₂O₂generating system, we found that about 80% of i-Tg was degraded and 24% of its iodine content was released. Our data point to a role for i-Tg in iodine storage and the involvement of TPO in i-Tg degradation and iodide release.