In vitro estimation of the rate of hexose phosphorylation, by sequential pulsing with [3H]- and [14C]2-deoxy-D-glucose.
The rate of phosphorylation of 2-deoxy-D-glucose (2dGlc) was determined by incubating Schistosoma mansoni in vitro in [3H]2-deoxy-D-glucose; 60 sec after exposure to the [3H]dGlc, [14C]dGlc was added to the medium, and metabolic activity was arrested at 2 min by immersion of the tissue in ice-cold silicone oil. Column chromatographic separation of the neutral [3H]- and [14C]dGlc from the [3H]- and [14C]2-deoxy-D-glucose-6-phosphate permitted estimation of the quantity of [3H]dGlc phosphorylated in 2 min, and the proportion of [14C]dGlc phosphorylated in 1 min; thus a phosphorylation rate was determined from a single tissue sample. In male schistosomes derived from mouse infections 4.4 +/- 0.8% of the dGlc was phosphorylated each minute, and 4.2 +/- 0.9% in the females. Lower rates of phosphorylation were measured in schistosomes taken from hamsters where males phosphorylated 2.4 +/- 1.1% of the dGlc each minute, and in females 2.7 +/- 1.0%. These studies suggest the high rate of hexose utilization by schistosomes compares to the conscious rat brain, where 11% of the dGlc is phosphorylated each minute.
D000818:Animals; D002250:Carbon Radioisotopes; D006224:Cricetinae; D003837:Deoxy Sugars / Q000378:metabolism*; D003847:Deoxyglucose / Q000378:metabolism*; D005260:Female; D006600:Hexosephosphates / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D010766:Phosphorylation; D012550:Schistosoma mansoni / Q000378:metabolism; D014316:Tritium
D002250:Carbon Radioisotopes; D003837:Deoxy Sugars; D006600:Hexosephosphates; D014316:Tritium; D003847:Deoxyglucose