Effect of therapeutic concentration of lithium on live HEK293 cells; increase of Na+/K+-ATPase, change of overall protein composition and alteration of surface layer of plasma membrane
Jazyk angličtina Země Nizozemsko Médium print-electronic
Typ dokumentu časopisecké články, práce podpořená grantem
PubMed
28215700
DOI
10.1016/j.bbagen.2017.02.011
PII: S0304-4165(17)30055-7
Knihovny.cz E-zdroje
- Klíčová slova
- HEK293 cells, Lithium, Membrane biophysics, Na(+)/K(+)-ATPase, Proteomic analysis,
- MeSH
- buněčná membrána účinky léků metabolismus MeSH
- buněčné linie MeSH
- cytosol účinky léků metabolismus MeSH
- energetický metabolismus účinky léků MeSH
- HEK293 buňky MeSH
- lidé MeSH
- lithium farmakologie MeSH
- mitochondrie účinky léků metabolismus MeSH
- ouabain farmakologie MeSH
- proteomika metody MeSH
- sodíko-draslíková ATPasa metabolismus MeSH
- upregulace účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- lithium MeSH
- ouabain MeSH
- sodíko-draslíková ATPasa MeSH
BACKGROUND: The effect of long-term exposure of live cells to lithium cations (Li) was studied in HEK293 cells cultivated in the presence of 1mM LiCl for 7 or 21days. The alteration of Na+/K+-ATPase level, protein composition and biophysical state of plasma membrane was determined with the aim to characterize the physiological state of Li-treated cells. METHODS: Na+/K+-ATPase level was determined by [3H]ouabain binding and immunoblot assays. Overall protein composition was determined by 2D electrophoresis followed by proteomic analysis by MALDI-TOF MS/MS and LFQ. Li interaction with plasma membrane was characterized by fluorescent probes DPH, TMA-DPH and Laurdan. RESULTS: Na+/K+-ATPase was increased in plasma membranes isolated from cells exposed to Li. Identification of Li-altered proteins by 2D electrophoresis, MALDI-TOF MS/MS and LFQ suggests a change of energy metabolism in mitochondria and cytosol and alteration of cell homeostasis of calcium. Measurement of Laurdan generalized polarization indicated a significant alteration of surface layer of isolated plasma membranes prepared from both types of Li-treated cells. CONCLUSIONS: Prolonged exposure of HEK293 cells to 1mM LiCl results in up-regulation of Na+/K+-ATPase expression, reorganization of overall cellular metabolism and alteration of the surface layer/polar head-group region of isolated plasma membranes. GENERAL SIGNIFICANCE: Our findings demonstrate adaptation of live HEK293 cell metabolism to prolonged exposure to therapeutic concentration of Li manifested as up-regulation of Na+/K+-ATPase expression, alteration of protein composition and change of the surface layer of plasma membrane.
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