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Micelles of poly(oxyethylene)-poly(oxypropylene) block copolymer (pluronic) as a tool for low-molecular compound delivery into a cell: phosphorylation of intracellular proteins with micelle incorporated [gamma-32P]ATP.
Authors:V I Slepnev  L E Kuznetsova  A N Gubin  E V Batrakova  Kabanov" target="_blank">A V Alakhov VYuKabanov
Institution:Department of Bioregulation, Research Center of Molecular Diagnostics and Therapy, Moscow, Russia.
Abstract:Pluronic P85 (poly(oxyethylene)-poly(oxypropylene) block copolymer) was used for in vitro delivery of gamma-32P]ATP into intact Jurkat cells. Negatively charged ATP molecules are not able to penetrate cell plasma membrane. Hence, exogenous gamma-32P]ATP added to a cell culture does not participate in phosphorylation of intracellular proteins. The addition to cells of gamma-32P]ATP solubilized in positively charged (containing dodecylamine) pluronic micelles results in considerable increase of protein phosphorylation. In this case the treatment of intact cells with alkaline phosphatase (resulting in dephosphorylation of external proteins) causes no essential decrease of 32P]-incorporation in cell proteins. That gives an evidence of delivery of solubilized ATP into a cell. Under the experimental conditions used, pluronic micelles neither influence the viability of cells nor permeabilize cell plasma membrane.
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