Increase in the affinity of the uridine phosphorylation system for ATP after serum or insulin activation of 3T3 fibroblasts.

Abstract

The stimulation of uridine uptake, brought about by the addition of serum or insulin to quiescent 3T3 fibroblasts, is associated in the half-saturation concentration of the uridine phosphorylating system for the substrate ATP, with relatively little change in the maximum uptake or in the affinity for uridine. In stimulated cells the Km towards ATP fell in the range 0.053--0.187 mM, while V max was 34 to 52 pmoles/10(6) cells/min. In quiescent cells these values were 2.89--4.22 mM and 74.5--126 pmoles/10(6) cells/min, respectively. No difference was found, however, between the Km's for ATP when phosphorylation of uridine was determined using cell-free extracts prepared from either quiescent cells or from stimulated cells.