Use of digitonin fractionation to determine mitochondrial transmembrane ion distribution in cells during anoxia.
A method to determine intracellular distribution of ions and metabolites under conditions of low oxygen concentration was developed. The technique involves rapid digitonin fractionation and addition of cyanide to inhibit reoxygenation. Applicability of the procedure was examined by studying the distribution of the lipophilic ion triphenylmethylphosphonium, the weak acid 5,5-dimethyloxazolidine-2,4-dione, and adenine nucleotides.
D000227:Adenine Nucleotides / Q000378:metabolism; D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D004072:Digitonin; D004114:Dimethadione / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D000860:Hypoxia / Q000378:metabolism*; D007477:Ions / Q000378:metabolism*; D008297:Male; D008564:Membrane Potentials; D008930:Mitochondria, Liver / Q000378:metabolism*; D009861:Onium Compounds / Q000378:metabolism; D010101:Oxygen Consumption; D051381:Rats; D014320:Triphenylmethyl Compounds / Q000378:metabolism
D000227:Adenine Nucleotides; D007477:Ions; D009861:Onium Compounds; D014320:Triphenylmethyl Compounds; C011561:triphenylmethylphosphonium; D004114:Dimethadione; D004072:Digitonin