Determination of intracellular pH and compartmentation using diffusion-weighted NMR spectroscopy with pH-sensitive indicators.
The intracellular pH (pHi) of a series of cancer cell lines was determined using the pH-sensitive indicators imidazole (Im) or histidine (His) and diffusion-weighted (DW) proton NMR spectroscopy. The DW method allows the observation at high magnetic field gradient values of only the slow-moving (intracellular) components, thus ensuring complete separation between intra- and extracellular components. Using the chemical shift difference (deltadelta) between the imidazole ring C2-H and C4(5)-H peaks, we were able to measure the pHi independently of chemical shift standardization. With His, the cell lines gave pHi values of approximately 6.5-7.0, whereas with Im, a second, more acidic compartment (pHi = 5.5-5.8) was also observed. An inverse correlation was also found between pHi and the intracellular lactate concentration. This method may be applicable to in vivo pH determinations.
Cohen
Jack S
JS
Department of Pharmacology, Faculty of Medicine, Hebrew University, Jerusalem, Israel.
Motiei
Menachem
M
Carmi
Shani
S
Shiperto
Dana
D
Yefet
Odeya
O
Ringel
Israel
I
D002138:Calibration; D002451:Cell Compartmentation; D045744:Cell Line, Tumor; D006639:Histidine / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007093:Imidazoles / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*
D007093:Imidazoles; D006639:Histidine; C029899:imidazole