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Identification of thyroglobulin domain(s) involved in cell-surface binding and endocytosis.

Abstract : Thyroglobulin (Tg) binds to cell surfaces through various binding sites of high, moderate and low affinity. We have previously shown that binding with low to moderate affinity is pH dependent, selective, but not tissue specific. To identify the regions of Tg involved in this cell surface binding, we studied the binding of (125)I-labeled cyanogen bromide peptides from human Tg to cell surfaces of thyroid cells (inside-out follicles) and of CHO cells. Electrophoretic analysis of cell homogenates after binding of native or of reduced and alkylated (125)I-labeled peptides showed that three peptides, P1, P2 and P3, were always associated with the cells. Sequence analysis allowed the identification of P1 (Ser-2445 to Met-2596 or Met-2610) and P2 (Phe-2156 to Met-2306). P3 proved to be a mixture of several peptides among which two were identified: P3-1 (Cys-1306 to Met-1640) and P3-2 (Cys-2035 to Met-2413) which includes P2. P1, P2 and P3-2 are entirely (P1) or partly (P2 and P3-2) located in the C-terminal domain of Tg homologous with acetylcholinesterase. The smallest peptides, P1 and P2, were purified by preparative electrophoresis. They both displayed strong binding properties towards cell surfaces. Inhibition experiments of (125)I-labeled Tg binding by P1 or P2 indicated that they were involved in Tg binding to cell surfaces. All the other peptides tested for their binding abilities were either not or only poorly involved in Tg binding to cell surfaces, which suggested that P1 and P2 are major Tg sites of binding to cell surfaces. These two peptides are not involved in the binding of Tg to the known Tg 'receptors' described in the literature, to which recycling, transcytosis and regulation functions have been ascribed. Thus they are potential tools to identify cell surface components involved in the process of Tg endocytosis leading to lysosomal degradation.
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https://hal.archives-ouvertes.fr/hal-02107434
Contributor : Jean-Louis Franc <>
Submitted on : Tuesday, April 23, 2019 - 4:28:53 PM
Last modification on : Wednesday, April 24, 2019 - 1:38:10 AM

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  • HAL Id : hal-02107434, version 1
  • PUBMED : 11431154

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Sandrine Siffroi-Fernandez, Frédéric Delom, Mc Nlend, J Lanet, Jean-Louis Franc, et al.. Identification of thyroglobulin domain(s) involved in cell-surface binding and endocytosis.. Journal of Endocrinology, BioScientifica, 2001, 170 (1), pp.217-26. ⟨hal-02107434⟩

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