The cascade reaction of tyrosine phosphorylation provides an attractive mechanism to regulate the metabolism and growth of normal and transformed cells. Thus, considerable effort has been made to identify the cellular...The cascade reaction of tyrosine phosphorylation provides an attractive mechanism to regulate the metabolism and growth of normal and transformed cells. Thus, considerable effort has been made to identify the cellular substrates of the tyrosine kinases. In this work, the activity of the Insulin receptor tyrosine kinase and endogenous substrate in transformed lymphocytes were studied. Purified human T lymphocytes incubated with phytohemagglutinin (PHA) for 72 hours served as transformed cells, when labeled with [32P] - orthophosphate it appeared that the insulin-dependent protein kinase activity in the transformed cells increased 9- fold. In search for the physiologically significant substrates by using polyclonal antiphosphotyrosine antibody to immunoprecipitate phosphotyrosine-containing proteins that produced in the intact cell during insulin stimulation, a protein with molecular weight of 45kDa as identified and designated as PP45, which occurred during the 5 minutes response of lymphocytes to insulin.展开更多
文摘The cascade reaction of tyrosine phosphorylation provides an attractive mechanism to regulate the metabolism and growth of normal and transformed cells. Thus, considerable effort has been made to identify the cellular substrates of the tyrosine kinases. In this work, the activity of the Insulin receptor tyrosine kinase and endogenous substrate in transformed lymphocytes were studied. Purified human T lymphocytes incubated with phytohemagglutinin (PHA) for 72 hours served as transformed cells, when labeled with [32P] - orthophosphate it appeared that the insulin-dependent protein kinase activity in the transformed cells increased 9- fold. In search for the physiologically significant substrates by using polyclonal antiphosphotyrosine antibody to immunoprecipitate phosphotyrosine-containing proteins that produced in the intact cell during insulin stimulation, a protein with molecular weight of 45kDa as identified and designated as PP45, which occurred during the 5 minutes response of lymphocytes to insulin.
