Determinants for chromogranin A sorting into the regulated secretory pathway are also sufficient to generate granule-like structures in non-endocrine cells
| Autor: | Hansruedi Stettler, Martin Spiess, Bérengère Fayard, Cristina Prescianotto-Baschong, Nicole Beuret, Laurent Taupenot |
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| Rok vydání: | 2009 |
| Předmět: |
Enteroendocrine cell
Biochemistry Epitope Cell Line Green fluorescent protein Epitopes Mice Chlorocebus aethiops Animals Microscopy Immunoelectron Molecular Biology Secretory pathway Secretory Pathway biology Secretory Vesicles Granule (cell biology) Chromogranin A Cell Biology Recombinant Proteins Rats Cell biology Secretory protein Cell culture biology.protein Endocrine Cells Biomarkers Gene Deletion |
| Zdroj: | Biochemical Journal. 418:81-91 |
| ISSN: | 1470-8728 0264-6021 |
| Popis: | In endocrine cells, prohormones and granins are segregated in the TGN (trans-Golgi network) from constitutively secreted proteins, stored in concentrated form in dense-core secretory granules, and released in a regulated manner on specific stimulation. The mechanism of granule formation is only partially understood. Expression of regulated secretory proteins, both peptide hormone precursors and granins, had been found to be sufficient to generate structures that resemble secretory granules in the background of constitutively secreting, non-endocrine cells. To identify which segment of CgA (chromogranin A) is important to induce the formation of such granule-like structures, a series of deletion constructs fused to either GFP (green fluorescent protein) or a short epitope tag was expressed in COS-1 fibroblast cells and analysed by fluorescence and electron microscopy and pulse-chase labelling. Full-length CgA as well as deletion constructs containing the N-terminal 77 residues generated granule-like structures in the cell periphery that co-localized with co-expressed SgII (secretogranin II). These are essentially the same segments of the protein that were previously shown to be required for granule sorting in wild-type PC12 (pheochromocytoma cells) cells and for rescuing a regulated secretory pathway in A35C cells, a variant PC12 line deficient in granule formation. The results support the notion that self-aggregation is at the core of granule formation and sorting into the regulated pathway. |
| Databáze: | OpenAIRE |
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