Ion-selective electrode for transmembrane pH difference measurements.
A triethylammonium-sensitive electrode was constructed using sodium tetrakis[3,5-bis(2-methoxyhexafluoro-2-propyl)phenyl]borate as an ion-exchanger and benzyl 2-nitrophenyl ether as a solvent mediator in a poly(vinylchloride) membrane matrix and was used to determine the pH difference across a cell membrane. The method is based on monitoring of the pH gradient-induced uptake of triethylammonium in situ. The triethylammonium electrode exhibited a near-Nernstian response to triethylammonium in the concentration range of 5 x 10(-6)-1 x 10(-2) M with a slope of 58.5 mV per concentration decade in a buffer solution composed of 150 mM NaCl and 10 mM NaH2PO4/Na2HPO4 (pH 7.5). The limit of detection was 1 microM. In experiments using liposomes, the uptake of triethylammonium into liposomes was quantitatively induced according to the pH difference across the liposomal membrane. The transmembrane pH differences in Escherichia coli cells and the light-induced pH differences across the envelope vesicles of Halobacterium halobium were successfully determined by the present method.
Katsu
T
T
Faculty of Pharmaceutical Sciences, Okayama University, Tsushima, Okayama 700-8530, Japan. katsu@pheasant.pharm.okayama-u.ac.jp
Nakagawa
H
H
Kanamori
T
T
Kamo
N
N
Tsuchiya
T
T
D002462:Cell Membrane / Q000737:chemistry*; D004926:Escherichia coli / Q000737:chemistry; D006217:Halobacterium / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008081:Liposomes; D008839:Microelectrodes
D008081:Liposomes