SERCaMP: a carboxy-terminal protein modification that enables monitoring of ER calcium homeostasis
| Autor: | Mark J. Henderson, Emily S. Wires, Christopher T. Richie, Kathleen A. Trychta, Brandon K. Harvey |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
Male
Programmed cell death Receptors Peptide Recombinant Fusion Proteins Green Fluorescent Proteins Primary Cell Culture chemistry.chemical_element Biology Calcium Endoplasmic Reticulum Protein Engineering Sarcoplasmic Reticulum Calcium-Transporting ATPases Rats Sprague-Dawley Transduction Genetic Caffeine Methods Animals Homeostasis Humans Calcium Signaling Luciferases Molecular Biology Calcium signaling Neurons Calcium metabolism Endoplasmic reticulum HEK 293 cells Articles Cell Biology Dependovirus Endoplasmic Reticulum Stress Rats Transport protein Protein Transport HEK293 Cells Liver Biochemistry chemistry Central Nervous System Stimulants |
| Zdroj: | Molecular Biology of the Cell |
| ISSN: | 1939-4586 1059-1524 |
| Popis: | Depletion of ER calcium can lead to cell death and is implicated in numerous diseases. Secreted ER calcium-monitoring proteins (SERCaMPs) are secreted in response to ER calcium depletion. SERCaMPs identified ER calcium depletion in primary neurons exposed to glutamate, hyperthermia, and coxibs and in rat liver after a single exposure to thapsigargin. Endoplasmic reticulum (ER) calcium homeostasis is disrupted in diverse pathologies, including neurodegeneration, cardiovascular diseases, and diabetes. Temporally defining calcium dysregulation during disease progression, however, has been challenging. Here we describe secreted ER calcium-monitoring proteins (SERCaMPs), which allow for longitudinal monitoring of ER calcium homeostasis. We identified a carboxy-terminal modification that is sufficient to confer release of a protein specifically in response to ER calcium depletion. A Gaussia luciferase (GLuc)–based SERCaMP provides a simple and sensitive method to monitor ER calcium homeostasis in vitro or in vivo by analyzing culture medium or blood. GLuc-SERCaMPs revealed ER calcium depletion in rat primary neurons exposed to various ER stressors. In vivo, ER calcium disruption in rat liver was monitored over several days by repeated sampling of blood. Our results suggest that SERCaMPs will have broad applications for the long-term monitoring of ER calcium homeostasis and the development of therapeutic approaches to counteract ER calcium dysregulation. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|