Dexamethasone inhibits sugar-dependent phorbol myristate acetate (PMA)-stimulated superoxide production and 2-deoxy-D-glucose (2-dGlc) transport in rat peritoneal macrophages (Rist, R.J., Jones, G.E. and Naftalin, R.J. (1991) Biochem. J. 278, 119-128; Rist, R.J. and Naftalin, R.J. (1991) Biochem J. 278, 129-135). Here it is shown that with glucose as a substrate, dexamethasone (0.1 microM) acts as a non-competitive inhibitor of PMA-induced superoxide production; decreasing the maximal rate of superoxide production (P < 0.001) without altering the Km. In contrast, with 2-dGlc as a substrate, dexamethasone shows competitive inhibition of PMA-stimulated superoxide production; increasing the Km of superoxide production, (P < 0.001) without altering the Vmax. The maximal rate of PMA-stimulated superoxide production with glucose as substrate is 10-12-fold in excess of the maximal rate with 2-dGlc as substrate. Diphenylene iodonium (DPI) is a non-competitive inhibitor of PMA-stimulated glucose-dependent superoxide production in macrophages, (Ki = 1-5 microM) and significantly reduces the activity of the PMA-induced hexose monophosphate shunt, (HMPS) (P < 0.01). However, DPI (1 microM) has no significant effect on the PMA-induced increase in 2-dGlc uptake, suggesting that the stimulus for HMPS activity and superoxide production is separate from the stimulus for hexose transport. A model is described which explains the observed differences in hexose transport and glucose- and 2-dGlc-dependent superoxide production in terms of the differences in metabolism of the two sugars. Accumulation of free 2-dGlc within the cytosol leads to saturation of hexokinase and hence, the effects of PMA and dexamethasone, which alter the coupling between hexokinase and the transporter, are only observed at low concentrations of 2-dGlc, where it is accumulated to sub-saturating levels. Since glucose is completely metabolized within the cell, PMA and dexamethasone increase and decrease, respectively, net uptake of sugar and superoxide production at all glucose concentrations.