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

0474133 - FGÚ 2018 RIV NL eng J - Článek v odborném periodiku
Vošahlíková, Miroslava - Ujčíková, Hana - Chernyavskiy, Oleksandr - Brejchová, Jana - Roubalová, Lenka - Alda, M. - Svoboda, Petr
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.
Biochimica et Biophysica Acta-General Subjects. Roč. 1861, č. 5 (2017), s. 1099-1112. ISSN 0304-4165. E-ISSN 1872-8006
Grant CEP: GA ČR(CZ) GAP207/12/0919; GA ČR(CZ) GA15-16605S; GA ČR(CZ) GA17-07070S
Institucionální podpora: RVO:67985823
Klíčová slova: lithium * HEK293 cells * Na+/K+-ATPase * membrane biophysics * proteomic analysis
Obor OECD: Biochemistry and molecular biology
Impakt faktor: 3.679, rok: 2017

The effect of long-term exposure of live cells to lithium cations (Li) was studied in HEK293 cells cultivated in the presence of 1 mM LiCl for 7 or 21 days. 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. 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. 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. Prolonged exposure of HEK293 cells to 1 mM 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. 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.
Trvalý link: http://hdl.handle.net/11104/0271245