Detection and quantification of the vacuolar H+ATPase using theLegionellaeffector protein SidK
| العنوان: | Detection and quantification of the vacuolar H+ATPase using theLegionellaeffector protein SidK |
|---|---|
| المؤلفون: | Michelle E. Maxson, Yazan M. Abbas, Jing Ze Wu, Jonathan D. Plumb, Sergio Grinstein, John L. Rubinstein |
| المصدر: | Journal of Cell Biology. 221 |
| بيانات النشر: | Rockefeller University Press, 2022. |
| سنة النشر: | 2022 |
| مصطلحات موضوعية: | Vacuolar Proton-Translocating ATPases, Legionella, Saccharomyces cerevisiae, Cell Biology, Hydrogen-Ion Concentration, Fluorescence, Rats, Mice, RAW 264.7 Cells, Bacterial Proteins, Phagosomes, Animals, Humans, Lysosomes, HeLa Cells |
| الوصف: | Acidification of secretory and endocytic organelles is required for proper receptor recycling, membrane traffic, protein degradation, and solute transport. Proton-pumping vacuolar H+ ATPases (V-ATPases) are responsible for this luminal acidification, which increases progressively as secretory and endocytic vesicles mature. An increasing density of V-ATPase complexes is thought to account for the gradual decrease in pH, but available reagents have not been sufficiently sensitive or specific to test this hypothesis. We introduce a new probe to localize and quantify V-ATPases. The probe is derived from SidK, a Legionella pneumophila effector protein that binds to the V-ATPase A subunit. We generated plasmids encoding fluorescent chimeras of SidK1-278, and labeled recombinant SidK1-278 with Alexa Fluor 568 to visualize and quantify V-ATPases with high specificity in live and fixed cells, respectively. We show that V-ATPases are acquired progressively during phagosome maturation, that they distribute in discrete membrane subdomains, and that their density in lysosomes depends on their subcellular localization. |
| تدمد: | 1540-8140 0021-9525 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::e90aaed18dcc8282e0cbaf9016131633 https://doi.org/10.1083/jcb.202107174 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....e90aaed18dcc8282e0cbaf9016131633 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15408140 00219525 |
|---|