Differential effect of vanadate on receptor-mediated endocytosis of the asialoglycoprotein receptor in hepatocytes from normal and diabetic rats.

Insulin-dependent diabetes has been shown to affect several aspects of receptor-mediated endocytosis. Since vanadate, a phosphate analogue, is known to exert insulin-like actions in target tissues, we studied the effects of vanadate on the endocytosis of the asialoglycoprotein receptor (ASGP-R) after its administration either in vivo (oral therapy) and/or in vitro by direct incubation of isolated hepatocytes with vanadate. The surface binding, internalization, and degradation of 3H-asialoorosomucoid (3H-ASOR), a prototype ligand of the ASGP-R, were decreased in diabetic rats by approximately 36.

Five isoenzymes of protein kinase C are expressed in normal and STZ-diabetic rat hepatocytes: effect of phorbol 12-myristate 13-acetate.

Using isoenzyme-specific antisera, five Protein Kinase Cs (PKCs) were detected in cytosol and membrane hepatocytes from normal rats: PKC alpha (80 kDa), PKC beta II (40, 50, 55, 85 kDa), PKC delta (74, 76 kDa), PKC epsilon (95 kDa), PKC zeta (65, 70 kDa). STZ-diabetes induced a lower expression of the five PKCs, a higher localization in the cytosol, a preferential expression of PKC delta as the 76 kDa phosphorylated species and a decreased kinase activity towards Histone III-S. A 1 microM phorbol 12-myristate 13-acetate (PMA) incubation induced similar translocation to the membrane of PKCs alpha, native 85 kDa beta II and epsilon.

Differential expression of five protein kinase C isoenzymes in FAO and HepG2 hepatoma cell lines compared with normal rat hepatocytes.

We analyzed the expression of five protein kinase C (PKC) isoforms in cytosolic and membrane fractions from normal rat hepatocytes compared with those of two tumorigenic cell lines FAO and HepG2. Western blots with PKC-specific isoenzymes polyclonal antibodies provide evidences for the presence of the five isoforms alpha, beta II, delta, epsilon and zeta in normal rat hepatocytes. In hepatoma cells, we show differences in the level of expression, the molecular sizes and the responses to Phorbol 12-myristate 13-acetate (PMA).

Activation of phosphotyrosine phosphatase activity is associated with decreased differentiation in adult bovine lens.

The postnatal vertebrate eye lens provides an opportunity to study possible involvement of reversible protein phosphorylation in the differentiation process of epithelial cells. Epithelial cells at the lens equator, indeed, differentiate continuously into fiber cells throughout life but this capacity progressively decreases with age. Here we describe the characterization of a phosphotyrosine-protein phosphatase(s) (PTPase(s)) in the equatorial epithelium of bovine lens which exhibits a high level of specific activity.

A high phosphotyrosine phosphatase activity is present in differentiating bovine lens cells.

Activity of phosphotyrosine-protein phosphatases (PTPases) has been investigated in the different cellular regions of bovine eye lens. PTPases were tested in cellular detergent extracts using phospholabelled synthetic peptides and p-nitrophenyl phosphate. We show that a high PTPase activity is only present in cells which undergo differentiation, namely the equatorial epithelium and cortex fiber cells.