Alteration of rat liver endoplasmic reticulum Ca(2+)-ATPase thiol integrity by ciprofibrate, a peroxisome...

Ciprofibrate (CP), a peroxisome proliferator, has been shown to reduce rat liver endoplasmic reticulum (ER) Ca(2+)-ATPase activity both in vitro and in vivo. The ER Ca(2+)-ATPase is highly susceptible to thiol reactivity, and maintenance of maximal enzyme activity is critically dependent upon the integrity of these thiol groups. We therefore investigated whether CP alters ER Ca(2+)-ATPase thiol groups as a possible mechanism of enzyme inhibition. Using a thiol immunoblot technique, free thiol groups specifically on the ER Ca(2+)-ATPase were localized. Exposure of freshly isolated rat liver microsomes to CP (500 microM) resulted in a loss of sulfhydryl reactivity on the ER Ca(2+)-ATPase protein at 107 kDa, as identified using the thiol immunoblot assay. However, when rat liver microsomes were exposed to CP in the presence of reduced glutathione (GSH), thiol groups on the ER Ca(2+)-ATPase were protected. Also, the reduction of ER Ca(2+)-ATPase activity by CP could be ameliorated by co-incubation of rat liver microsomes with GSH. These observations indicate that CP reduces rat liver ER Ca(2+)-ATPase activity through interactions with free thiol groups located on this enzyme.