DEPLETION OF RED-CELL ATP BY INCUBATION WITH 2-DEOXYGLUCOSE

  • 1 January 1979
    • journal article
    • research article
    • Vol. 94  (4) , 617-623
Abstract
To devise a more physiologic system for measuring depletion of red cell [RBC] ATP levels, the effect of incubating human erythrocytes with 2-deoxyglucose was investigated. ATP depletion proceeded very slowly at a 10 mM concentration of 2-deoxyglucose, a level exceeding the Km of hexokinase for this substrate by more than 10-fold. At 160 mM concentration of 2-deoxyglucose, ATP depletion proceeded rapidly enough that nearly 90% of ATP disappeared from the RBC after 2 1/2 h of incubation. To explain this observation, additional studies were carried out. 2-Deoxyglucose penetrated rapidly into RBC. Phosphorylation of 2-deoxyglucose in RBC was inhibited by both products of the 2-deoxyglucose-phosphorylating reaction, i.e., 2-deoxyglucose-6-phosphate and ADP. Inhibition of 2-deoxyglucose phosphorylation diminished at higher-than-physiologic pH levels. RBC may be relatively rapidly depleted of ATP by incubation with 100 mM 2-deoxyglucose in a saline-phospate-buffered medium, pH 7.8. In such rapidly depleted cells, morphologic changes formerly attributed to ATP depletion, do not occur.