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Biochim Biophys Acta Gen Subj. 2017 May;1861(5 Pt A):1099-1112. doi: 10.1016/j.bbagen.2017.02.011. Epub 2017 Feb 13.

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.

Author information

1
Department of Biomathematics, Institute of Physiology of the Czech Academy of Sciences, Prague, Czech Republic. Electronic address: miroslava.vosahlikova@fgu.cas.cz.
2
Department of Biomathematics, Institute of Physiology of the Czech Academy of Sciences, Prague, Czech Republic.
3
Department of Psychiatry, Dalhousie University, Halifax, Nova Scotia, Canada; National Institute of Mental Health, Klecany, Czech Republic.

Abstract

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.

KEYWORDS:

HEK293 cells; Lithium; Membrane biophysics; Na(+)/K(+)-ATPase; Proteomic analysis

PMID:
28215700
DOI:
10.1016/j.bbagen.2017.02.011
[Indexed for MEDLINE]

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