Novel Cl(−)-dependent intracellular pH regulation in murine MDR 1 transfectants and potential implications Journal Article


Authors: Roepe, P. D.; Weisburg, J. H.; Luz, J. G.; Hoffman, M. M.; Wei, L. Y.
Article Title: Novel Cl(−)-dependent intracellular pH regulation in murine MDR 1 transfectants and potential implications
Abstract: Previously [Luz et al. (1994) Biochemistry 33, 7239–7249], we determined that Cl−- and −HCO3-dependent pHi homeostasis was perturbed in multidrug resistant (MDR) cells created by transfecting LR73 Chinese hamster ovary fibroblasts with wild-type mu (murine) MDR 1 (Gros et al., 1991). Via single-cell photometry experiments performed under various conditions, we are now able to separate Na+-dependent and Na+-independent components of Cl−/−HCO3 exchange in the MDR transfectants and the parental LR73 cells. Cl−-dependent, Na+-independent reacidification of pHi, mediated by the anion exchanger 2 isoform in LR73 cells, is dramatically inhibited by mild overexpression of MDR protein. Analysis of H+ flux at different pHj shows that Cl−-dependent reacidification approaches 0.2 mM H+/s for LR73 cells at pHi = 8.0 but is at least 10-fold slower for MDR 1 transfectants that were never exposed to chemotherapeutics (EX4N7 cells). MDR 1 transfectants selected on the chemotherapeutic vinblastine (1–1 cells), which express approximately 10-fold more MDR protein relative to EX4N7 cells, exhibit similar behavior; however, alterations in Cl−-dependent pH, regulation are more severe. Hypotonic conditions, which have been shown to increase anomalous Cl− conductance in some cells overexpressing MDR protein (Valverde et al., 1992), are found to amplify the altered pHi homeostasis features in the primary transfectants that express lower levels of MDR protein such that they then mimic the behavior of the drug-selected cells that express substantially more MDR protein. Verapamil reverses the anomalous behavior. In addition, removal of CO2 causes similar pHi changes in all cell types, whereas removal of −HCO3 in the presence of Cl− leads to greater changes in pHi for the MDR cells. Transfectants harboring mutant MDR protein that is unable to confer the MDR phenotype (K432R/K1074R mu MDR 1) perform Na+-independent Cl−/−HCO3 exchange similar to the parental LR73 cells. These data may help to further refine models for MDR protein function in MDR cells. © 1994, American Chemical Society. All rights reserved.
Keywords: nonhuman; animal cell; animal; mice; cells, cultured; gene expression; drug resistance; transfection; animalia; genetic transfection; membrane glycoproteins; carrier proteins; murinae; homeostasis; verapamil; proton transport; multidrug resistance; hydrogen-ion concentration; p-glycoprotein; cho cell; cho cells; cricetinae; cricetulus; cricetulus griseus; chlorides; chloride channel; cell ph; priority journal; article; support, non-u.s. gov't; support, u.s. gov't, p.h.s.; hamsters; chloride conductance
Journal Title: Biochemistry
Volume: 33
Issue: 36
ISSN: 0006-2960
Publisher: American Chemical Society  
Date Published: 1994-09-13
Start Page: 11008
End Page: 11015
Language: English
DOI: 10.1021/bi00202a021
PROVIDER: scopus c2 - 7916210
PUBMED: 7916210
DOI/URL:
Notes: Export Date: 14 January 2019 -- Article -- Source: Scopus
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  1. Paul D. Roepe
    26 Roepe