engleski

Effect of bisperoxovanadate-1,10-phenantroline on mitogen-activated protein (MAP) kinases in rat PC12 cells

Mitogen-activated protein (MAP) kinase cascades represent one of the major signal systems used by eukariotic cells to transduce extracellular signals into cellular responses. Activation of the MAP kinases plays an important role in many cellular processes such as cell proliferation, differentiation and cell death. Bisperoxovanadate-1,10-phenanthroline (bpV) is a strong insulin mimetic and potent protein-tyrosine-phosphatase inhibitor and activator of insulin receptor kinase. The effect of bpV on the level and activity of MAP kinases (p38, JNK 1, ERK 1 and ERK 2) in PC-12 cells (rat adrenal pheochromocytoma) was observed using different time of incubation and doses of bpV. To determine the levels of MAP kinases, cell extracts were fractionated by 12% SDS polyacrylamide gel electrophoresis, the separated proteins were probed with mentioned antibodys for MAP kinases. To examine kinase activity, the cell extracts were immunoprecipitated with antibodys for MAP kinases, separated and probed with anti-phospho-tyrosine antibody. The blots were determined using the enhanced chemiluminiscence reaction. The same steady-state level of p38, JNK 1, ERK 1 and ERK 2 were determined in control and treated cultures without regard to time of incubation and doses of bpV. The tyrosine phosphorylation of JNK 1, ERK 1 and ERK 2 was observed after 20 minutes, and of p38 after one houre, with 100 mM bpV.

Bisperoxovanadate-1;10-phenantroline; phosphotyrosine phosphatase inhibitor; MAP kinases

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