Amino acid transport in multidrug-resistant Chinese hamster ovary cells.
In the process of assessing the effect of anthracycline drugs on cellular membrane function in cultured multidrug resistant (MDR) and its parental cells, experiments were undertaken to investigate the kinetics of neutral amino acid membrane transport (the sodium dependent A and ASC systems). P-glycoprotein, a high molecular weight energy requiring integral membrane protein responsible for actively pumping drugs out of cells, has been shown to be overexpressed in MDR cells. It was our hypothesis that its presence might affect other membrane energy requiring systems such as amino acid transport. On establishing the concentrations of P-glycoprotein by western blotting in the two cell lines to be studied, the kinetics of membrane transport of the neutral amino acids alpha-aminoisobutyric acid (AIB) and serine (SER) were investigated using the CHRC5 multidrug resistant and AUX B1 parental Chinese hamster ovary (CHO) cells. In CHRC5 cells, the amount and rate (Vmax) of accumulated amino acids, was significantly depressed when compared to AUX B1 cells, however, there was no difference in the rates of amino acid efflux between these two cell lines. Using 1,6-diphenyl 1,3,5-hexatriene (DPH) polarization to evaluate the state of membrane fluidity in the two cell lines studied, it was seen that CHRC5 cells showed a slightly lower degree of polarization than that observed in AUX B1 cells. These results suggest, that the P-glycoprotein does not alter amino acid transport directly but may modify the activity or numbers of functional transport carriers.
Daly
S E
SE
Department of Biochemistry, University of Western Ontario, London, Canada.
Inch
R W
RW
Tustanoff
E R
ER
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D001693:Biological Transport, Active / Q000502:physiology; D015153:Blotting, Western; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism*; D006224:Cricetinae; D004161:Diphenylhexatriene / Q000737:chemistry; D004351:Drug Resistance / Q000502:physiology*; D005260:Female; D005454:Fluorescence Polarization; D008560:Membrane Fluidity / Q000502:physiology; D008562:Membrane Glycoproteins / Q000502:physiology*
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000596:Amino Acids; D008562:Membrane Glycoproteins; D004161:Diphenylhexatriene