Vicinal thiols are involved in inositol 1,2,3,5,6-pentakisphosphate 5-phosphatase activity from fetal calf thymus

Abstract

Inositol 1,2,3,5,6-pentakisphosphate (Ins(1,2,3,5,6)P₅) 5-phosphatase present in fetal calf thymus has been partially purified. This enzyme was inhibited dose-dependently by different thiol modifiers likeN-ethylmaleimide (NEM),p-chloromercuribenzene sulfonate (PCMBS), diamide, and phenylarsine oxide (PAO). The inhibition by PCMBS and diamide was protected by preincubation with dithiothreitol (DTT) and the phosphatase substrate, Ins(1,2,3,5,6)P₅. Diamide, a compound that specifically modifies vicinal thiol groups, also blocked the 5-phosphatase dose-dependently. Specificity of this blockade was proven by using dimercaptopropanol (DMP), a compound known to protect vicinal thiol groups. DMP prevented the enzyme from inhibition by diamide. These data suggest that vicinal thiols are involved in Ins(1,2,3,5,6)P₅5-phosphatase activity.