004 Datenverarbeitung; Informatik
Refine
Has Fulltext
- yes (5)
Is part of the Bibliography
- yes (5)
Year of publication
- 2015 (5) (remove)
Document Type
- Journal article (5)
Language
- English (5)
Keywords
- CD4+T cells (1)
- CD8+T cells (1)
- HHblits (1)
- I-tasser (1)
- III secretion (1)
- LC-MS/MS (1)
- V-antigen (1)
- WH2 domain (1)
- Yersinia enterocolitica (1)
- actin nucleation (1)
- apixaban (1)
- behavior (1)
- cell membranes (1)
- complex traits (1)
- cytokine profiling (1)
- diagnostic accuracy (1)
- direct oral anticoagulants (1)
- direct thrombin inhibitor (1)
- disruption project (1)
- electrolytes (1)
- factor XA inhibitor (1)
- flies (1)
- functional analysis (1)
- genetics (1)
- hypotonic (1)
- hypotonic solutions (1)
- isotonic (1)
- membrane proteins (1)
- memory immune responses (1)
- mice (1)
- natural variation (1)
- networks (1)
- nonhuman-primates (1)
- painful (1)
- performance liquid chromatography (1)
- permeability (1)
- pestis infection (1)
- pneumonic plague (1)
- quantification (1)
- recombinant protein rVE (1)
- resistance (1)
- serum (1)
- shootin-1 (1)
- spire (1)
- temperature (1)
- tonicity (1)
- vaccine (1)
- validation (1)
Institute
- Theodor-Boveri-Institut für Biowissenschaften (5) (remove)
Swelling-activated pathways for myo-inositol, one of the most abundant organic osmolytes in mammalian cells, have not yet been identified. The present study explores the SLC5A3 protein as a possible transporter of myo-inositol in hyponically swollen HEK293 cells. To address this issue, we examined the relationship between the hypotonicity-induced changes in plasma membrane permeability to myo-inositol Pino [m/s] and expression/localization of SLC5A3. Pino values were determined by cell volumetry over a wide tonicity range (100–275 mOsm) in myo-inositol-substituted solutions. While being negligible under mild hypotonicity (200–275 mOsm), Pino grew rapidly at osmolalities below 200 mOsm to reach a maximum of ∼3 nm/s at 100–125 mOsm, as indicated by fast cell swelling due to myo-inositol influx. The increase in Pino resulted most likely from the hypotonicity-mediated incorporation of cytosolic SLC5A3 into the plasma membrane, as revealed by confocal fluorescence microscopy of cells expressing EGFP-tagged SLC5A3 and super-resolution imaging of immunostained SLC5A3 by direct stochastic optical reconstruction microscopy (dSTORM). dSTORM in hypotonic cells revealed a surface density of membrane-associated SLC5A3 proteins of 200–2000 localizations/μm2. Assuming SLC5A3 to be the major path for myo-inositol, a turnover rate of 80–800 myo-inositol molecules per second for a single transporter protein was estimated from combined volumetric and dSTORM data. Hypotonic stress also caused a significant upregulation of SLC5A3 gene expression as detected by semiquantitative RT-PCR and Western blot analysis. In summary, our data provide first evidence for swelling-mediated activation of SLC5A3 thus suggesting a functional role of this transporter in hypotonic volume regulation of mammalian cells.