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Introduction of regulatory subunits of cAMP-dependent protein kinase type II in ZTZ cells with the use of erythrocyte ghosts
Authors:A G Aprikian  L Iu Zhuleva  M V Nesterova  E S Severin
Abstract:The dynamics of distribution of the regulatory subunit of cAMP-dependent protein kinase II following protein injection into 3T3 cells was studied. The cAMP-binding component of protein kinase was injected into the cells, using erythrocyte ghosts. The conditions for protein encapsulation into erythrocyte ghosts were elaborated. The optimal detergent concentrations, incubation time and conditions of vesicular closure following protein injection were selected. The above method provides for a high (50-55%) yield of the erythrocyte ghost-encapsulated protein with a minimum loss of enzymatic activity. Fusion of erythrocyte ghosts containing the labeled protein with 3T3 cells was carried out. Using the cytoradiography technique, the dynamics of distribution of the radiolabeled regulatory subunit within the cell was analyzed. It was demonstrated that after the regulatory subunit has reached the cytoplasm, the protein is translocated into the nucleus and is pooled there is the vicinity of the nucleoli.
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