Procathepsin V is secreted in a TSH regulated manner from human thyroid epithelial cells and is accessible to an activity-based probe
| Autor: | Sebastian Springer, Klaudia Brix, Matthew Bogyo, Dagmar Führer, Ekkehard Weber, Maren Rehders, Zeynep Hein, Alaa Al-Hashimi, Naphannop Sereesongsaeng, Christopher J. Scott, Roberta E. Burden, Vaishnavi Venugopalan |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Glycosylation
endocrine system diseases medicine.medical_treatment Medizin Thyroid Gland Fluorescent Antibody Technique Gene Expression Thyrotropin Endoplasmic Reticulum Cysteine cathepsins lcsh:Chemistry Cathepsin L Genes Reporter Thyroid stimulating hormone Cathepsin L2 Cathepsin V lcsh:QH301-705.5 Spectroscopy biology Chemistry Thyroid General Medicine Cell biology Computer Science Applications secretion Protein Transport medicine.anatomical_structure protein trafficking Thyroid function endocrine system Protein trafficking Article Catalysis Cell Line green fluorescent protein tagging Inorganic Chemistry medicine Humans Amino Acid Sequence Physical and Theoretical Chemistry Molecular Biology Secretion Thyroid Epithelial Cells Cathepsin cysteine cathepsins thyroid epithelial cells Cell Membrane Organic Chemistry Green fluorescent protein tagging thyroid stimulating hormone Cathepsins lcsh:Biology (General) lcsh:QD1-999 Thyroid epithelial cells biology.protein Thyroglobulin Lysosomes Biomarkers |
| Zdroj: | Al-Hashimi, A, Venugopalan, V, Rehders, M, Sereesongsaeng, N, Hein, Z, Springer, S, Weber, E, Führer, D, Bogyo, M S, Scott, C J, Burden, R E & Brix, K 2020, ' Procathepsin V is secreted in a TSH regulated manner from human thyroid epithelial cells and is accessible to an activity-based probe ', International Journal of Molecular Sciences, vol. 21, no. 23, 9140 . https://doi.org/10.3390/ijms21239140 International Journal of Molecular Sciences Volume 21 Issue 23 International Journal of Molecular Sciences, Vol 21, Iss 9140, p 9140 (2020) |
| Popis: | The significance of cysteine cathepsins for the liberation of thyroid hormones from the precursor thyroglobulin was previously shown by in vivo and in vitro studies. Cathepsin L is most important for thyroglobulin processing in mice. The present study aims at specifying the possible contribution of its closest relative, cysteine cathepsin L2/V, to thyroid function. Immunofluorescence analysis on normal human thyroid tissue revealed its predominant localization at the apical plasma membrane of thyrocytes and within the follicle lumen, indicating the secretion of cathepsin V and extracellular tasks rather than its acting within endo-lysosomes. To explore the trafficking pathways of cathepsin V in more detail, a chimeric protein consisting of human cathepsin V tagged with green fluorescent protein (GFP) was stably expressed in the Nthy-ori 3-1 thyroid epithelial cell line. Colocalization studies with compartment-specific markers and analyses of post-translational modifications revealed that the chimeric protein was sorted into the lumen of the endoplasmic reticulum and subsequently transported to the Golgi apparatus, while being N-glycosylated. Immunoblotting showed that the chimeric protein reached endo-lysosomes and it became secreted from the transduced cells. Astonishingly, thyroid stimulating hormone (TSH)-induced secretion of GFP-tagged cathepsin V occurred as the proform, suggesting that TSH upregulates its transport to the plasma membrane before it reaches endo-lysosomes for maturation. The proform of cathepsin V was found to be reactive with the activity-based probe DCG-04, suggesting that it possesses catalytic activity. We propose that TSH-stimulated secretion of procathepsin V is the default pathway in the thyroid to enable its contribution to thyroglobulin processing by extracellular means. |
| Databáze: | OpenAIRE |
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