1. We investigated the effect of the thiol reagent, thimerosal on calcium movements in the Jurkat T cell line. 2. Thimerosal induced a rise in cytosolic Ca2+ concentration due both to a release of Ca2+ from intracellular stores and a Ca2+ influx. 3. Thimerosal, released Ca2+ from the same intracellular stores than CD3 mAb and ionomycin. 4. Emptying the Ca2+ intracellular stores was accompanied by a marked decrease of phosphatidylserine synthesis indicating that phosphatidylserine synthesis occurs within or close to the endoplasmic reticulum Ca(2+)-stores as previously described in CD3-, ionomycin- or Ca(2+)-ATPase inhibitor-treated lymphocytes.
1. The effects of okadaic acid (OA) and phorbol-12-myristate-13-acetate (PMA) on protein phosphorylation were studied in human term placentas. 2. When samples treated with tumour promoters were compared with untreated samples, the phosphorylation of a 135 kDa protein was significantly decreased; OA also produced a decrease in phosphorylation of a 24 kDa protein. 3. Both substances produced an alteration in the proportions of bands of masses 170, 65 and 24 kDa, relative to total phosphorylation; PMA treatment also affected the band of mass 135 kDa. 4. Placental cell extracts were also subjected to Western blotting with a protein kinase C (PKC) antibody, reportedly specific for the alpha- and beta-isoforms. 5. Two immunoreactive proteins were detected; an 80 kDa band, presumably corresponding to the alpha- or beta-PKC, and a 64 kDa protein, which could be a degradation production of the 80 kDa protein or it could correspond to another form of the enzyme. The expression of PKC did not change on treatment with PMA.
1. High-Km aldose reductase purified from dog kidney inner medulla was easily converted into aldose reductase by incubation in the neutral buffer solution. 2. High-Km aldose reductase was found to be in multiple forms, and was separated into three kinds of species designated as a-, b- and c-forms by HPLC. 3. The a-form observed as a single peak by HPLC was assumed to be present in three forms (a1-, a2- and a3-forms), one was aldose reductase (a1-form) and the others were the precursors of aldose reductase (a2- and a3-form). 4. The b-form was rapidly converted into the a3-form, followed slowly by the a2-form and finally into the a1-form. 5. The c-form was either directly converted into the a1-form, or indirectly into the a2-form followed by the a1-form. 6. Four kinds of species (a2-, a3-, b- and c-forms) of high-Km aldose reductase were finally converted into aldose reductase (a1-form).
