Heterologous expression of mammalian Na/H antiporters in Saccharomyces cerevisiae

Biochimica Et Biophysica Acta
Hana FlegelovaHana Sychrova

Abstract

Na+/H+ antiporters, integral membrane proteins that exchange protons for alkali metal cations, play multiple roles in probably all living organisms (preventing cells from excessive amounts of alkali metal cations, regulating intracellular pH and cell volume). In this work, we studied the functionality of rat plasma membrane NHE1-3 exchangers upon their heterologous expression in alkali-metal-cation sensitive Saccharomyces cerevisiae, and searched for conditions that would increase their level in the plasma membrane and improve their functionality. Though three tested exchangers were partially localized to the plasma membrane (and two of them (NHE2 and NHE3) in an active form), the bulk of the synthesized proteins were arrested along the secretory pathway, mainly in the ER. To increase the level of exchangers in the yeast plasma membrane several approaches (truncation of C-terminal regulatory sequences, expression in mutant yeast strains, construction of rat/yeast protein chimeras, various growth conditions and chemical chaperones) were tested. The only increase in the amount of NHE exchangers in the plasma membrane was obtained upon expression in a strain with the npi1 mutation, which significantly lowers the level of Rsp5 ubiq...Continue Reading

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Citations

Jul 29, 2011·Planta·Vaishali MulangiPaul Morris
Aug 25, 2009·Journal of Structural and Functional Genomics·Erin QuartleyElizabeth J Grayhack
Aug 10, 2013·Analytical Biochemistry·Chengqiang WangXiaoming Bao
Mar 4, 2010·Microbiology and Molecular Biology Reviews : MMBR·Joaquín AriñoHana Sychrová
Apr 15, 2011·American Journal of Physiology. Renal Physiology·Alexander R KolbJeffrey L Brodsky
Jul 30, 2014·Applied Microbiology and Biotechnology·Anita EmmerstorferHarald Pichler
Sep 6, 2007·FEMS Microbiology Reviews·Areti Pantazopoulou, George Diallinas
Feb 16, 2010·Protein Expression and Purification·James LeungBernadette Byrne
Mar 30, 2007·American Journal of Physiology. Cell Physiology·Masafumi MatsushitaHiroshi Kanazawa
Dec 7, 2013·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Ines ArmandoPedro A Jose
Dec 14, 2007·The Journal of Biological Chemistry·Karine MoncoqHoward S Young
Aug 2, 2017·International Journal of Microbiology·Safaa A S Al-QaysiJamal Abd Al-Rahman Ibrahim
Jul 17, 2008·Molecular and Cellular Biochemistry·Heng Chen, Larry Fliegel

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