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Multivalent binding proteins, such as the yeast scaffold protein Sterile-5, coordinate the location of kinases by serving as platforms for the assembly of signaling units. Similarly, in mammalian cells the cyclic adenosine 3',5'-monophosphate-dependent protein kinase (PKA) and phosphatase 2B [calcineurin (CaN)] are complexed by an A kinase anchoring(More)
Previously, we showed caveolae contain a population of protein kinase C ␣ (PKC ␣) that appears to regulate membrane invagination. We now report that multiple PKC isoenzymes are enriched in caveolae of unstimulated fibroblasts. To understand the mechanism of PKC targeting, we prepared caveolae lacking PKC ␣ and measured the interaction of recombinant PKC ␣(More)
We have used immunocytofluorescence techniques to determine the subcellular distribution of the Ca2+, phospholipid-dependent protein kinase, protein kinase C (PKC). Using monoclonal antibodies that are specific for Type 3 (alpha) PKC, we have determined that there are least two pools of PKC in normal rat embryo fibroblasts (REF52 cells): diffuse cytoplasmic(More)
We have previously characterized specific binding of the phorbol ester tumor promoter [20-3H]phorbol 12,13-dibutyrate to several tissues, including mouse skin and brain. We report here that specific binding activity in chicken brain increases dramatically during development. In whole embryonic chicken brain, binding increased from 2.0 to 6.8 to 10.6 pmol/mg(More)
We have examined the immunocytochemical localization of protein kinase C (PKC) in NIH 3T3 cells using mAbs that recognize Type 3 PKC. In control cells, the immunofluorescent staining was similar with mAbs directed to either the catalytic or the regulatory domain of PKC. Type 3 PKC localized in a diffuse cytoplasmic pattern, while the nuclei were apparently(More)
Abstrad Treatment of HC1 1 mouse mammary epithelial cells with the lactogenic hormones dexamethasone, insulin, and proladin (DIP) leads to cellular differentiation and production of the milk protein 13-casein. The following experimental evidence suggests the involvement of protein kinase C (PKC) in DIP induced signal transdudion. Down-regulation of PKC by 1(More)
Metastasis requires cytoskeletal remodeling for migration, adhesion, and extravasation of metastatic cells. Although protein kinase C (PKC) is involved in tumor promotion/progression and cytoskeletal remodeling, its role in metastasis has not been defined. PKCdelta levels are increased in highly metastatic 13762NF mammary tumor cells (MTLn3) compared with(More)
Immunocytofluorescence studies were used to compare alpha-protein kinase C (PKC) localization in primary cultures of renal proximal tubule epithelial cells (RPTE) with E1A-immortalized and SV40-transformed derivatives. Both cytosolic and nuclear staining were apparent in all of the RPTE. In primary RPTE, Triton X-100-insoluble alpha-PKC was also apparent(More)
In quiescent cultures of 3T3 cells, plasminogen activator (PA) is found predominantly as a 75,000 dalton species. When quiescent cells are exposed to mitogenic agents such as phorbol myristate acetate, Ca++, or 25% serum, the absolute levels of PA in cell lysates may either increase or decrease. However, a consistent observation is that in the stimulated(More)
The role of individual protein kinase C (PKC) isoforms in progressive transformation of a rat embryo fibroblast cell line (REF52 cells) has been evaluated. Normal (REF A) cells were transfected with SV40 to produce the partially transformed REF B cell line. REF B cells are morphologically transformed but have only limited capacity for growth in soft agar.(More)