Calcium as a reliable marker for the quantitative assessment of endoplasmic reticulum stress in live cells
| Autor: | Paul F. Lebeau, Jae Hyun Byun, Richard C. Austin, Khrystyna Platko |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
TG
thapsigargin 0301 basic medicine Glycosylation SERCA Thapsigargin Glucose-regulated protein Endoplasmic Reticulum SERCA sarco/endoplasmic reticulum ATPase Biochemistry Cell Line ER endoplasmic reticulum 03 medical and health sciences chemistry.chemical_compound 4-PBA 4-phenylbutyrate UPR unfolded protein response GRP glucose-regulated protein TUDCA tauroursodeoxycholic acid TM tunicamycin Humans unfolded protein response (UPR) Molecular Biology calcium 030102 biochemistry & molecular biology biology Mag-Fluo-4 Endoplasmic reticulum Ca2+ calcium Hep G2 Cells Cell Biology Tunicamycin RyR ryanodine receptor Endoplasmic Reticulum Stress Cell biology HEK293 Cells 030104 developmental biology Microscopy Fluorescence chemistry CDN CDN 1163 Unfolded Protein Response biology.protein Unfolded protein response endoplasmic reticulum stress (ER stress) Chemical chaperone VP vasopressin Research Article |
| Zdroj: | The Journal of Biological Chemistry |
| ISSN: | 0021-9258 |
| DOI: | 10.1016/j.jbc.2021.100779 |
| Popis: | Calcium (Ca2+) is an essential mineral of endoplasmic reticulum (ER) luminal biochemistry because of the Ca2+ dependence of ER-resident chaperones charged with folding de novo proteins that transit this cellular compartment. ER Ca2+ depletion reduces the ability of chaperones to properly fold the proteins entering the ER, thus leading to an accumulation of misfolded proteins and the onset of a state known as ER stress. However, not all conditions that cause ER stress do so in a manner dependent on ER Ca2+ depletion. Agents such as tunicamycin inhibit the glycosylation of de novo polypeptides, a key step in the maturation process of newly synthesized proteins. Despite this established effect of tunicamycin, our understanding of how such conditions modulate ER Ca2+ levels is still limited. In the present study, we report that a variety of ER stress–inducing agents that have not been known to directly alter ER Ca2+ homeostasis can also cause a marked reduction in ER Ca2+ levels. Consistent with these observations, protecting against ER stress using small chemical chaperones, such as 4-phenylbutyrate and tauroursodeoxycholic acid, also attenuated ER Ca2+ depletion caused by these agents. We also describe a novel high-throughput and low-cost assay for the rapid quantification of ER stress using ER Ca2+ levels as a surrogate marker. This report builds on our understanding of ER Ca2+ levels in the context of ER stress and also provides the scientific community with a new, reliable tool to study this important cellular process in vitro. |
| Databáze: | OpenAIRE |
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