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.
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.
Slepnev
V I
VI
Department of Bioregulation, Research Center of Molecular Diagnostics and Therapy, Moscow, Russia.
Kuznetsova
L E
LE
Gubin
A N
AN
Batrakova
E V
EV
Alakhov VYu
Kabanov
A V
AV
D000255:Adenosine Triphosphate / Q000008:administration & dosage* / Q000378:metabolism; D002463:Cell Membrane Permeability; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D007962:Leukocytes / Q000378:metabolism; D008823:Micelles; D014677:Pharmaceutical Vehicles; D010766:Phosphorylation; D011060:Poloxalene; D011506:Proteins / Q000378:metabolism; D012995:Solubility; D014407:Tumor Cells, Cultured / Q000378:metabolism
D008823:Micelles; D014677:Pharmaceutical Vehicles; D011506:Proteins; D000255:Adenosine Triphosphate; D011060:Poloxalene