Biochemical assay of inhibitors of metabolic cooperation

A significant and reproducible enhancement of purine nucleotide synthesis from hypoxanthine occurs in HAT medium, when communication-competent hypoxanthine-guanine phosphoribosyltransferase (HGPRT+) cells are co-cultured with communication-competent (HGPRT-) LN cells. This enhancement of purine nucleotide synthesis is dependent upon the hypoxanthine concentration and upon the ratio of (HGPRT-): (HGPRT+) cells. Based upon these results a simple biochemical method for the detection of inhibitors of metabolic cooperation between (HGPRT+) cells and (HGPRT-) LN cells is presented. The biochemical method distinguishes inhibitors of metabolic cooperation from inhibitors of hypoxanthine uptake, of hypoxanthine phosphorylation and of nucleic acid synthesis, as well as from general metabolic inhibitors. This method has the advantage that it can be used on a relatively large number of cells, it is simple and not time-consuming, and distinguishes the inhibition of metabolic cooperation by compounds that have a variety of sites of inhibition.