Tyrosine kinase and phosphotyrosine phosphatase activity in human promyelocytic leukemia cells and human polymorphonuclear leukocytes.
Although an increase in protein phosphorylation on tyrosine was first noted as a result of cell transformation or the application of growth factors to cells, recent reports have shown high levels of tyrosine kinases in nondividing tissues. For that reason, we have investigated whether normal human polymorphonuclear leukocytes (PMN) contain tyrosine kinase and phosphatase activity. Using a copolymer of glutamine: tyrosine as a substrate for the phosphotransferase reaction, we have demonstrated that PMN contain a cytosolic tyrosine kinase activity that elutes as a single peak from Sephacryl S-200 chromatography and has a molecular weight of 70 kilodaltons. Human promyelocytic leukemia cells (HL-60), contain a similar activity (as demonstrated by column chromatography), with only 25% of the activity found in PMN. This cytosolic tyrosine kinase can phosphorylate angiotensin II and a fragment of the src protein containing tyrosine 416, which suggests a similar substrate specificity to other tyrosine-phosphorylating protein kinases. In addition, we have demonstrated that PMN have double the amount of phosphotyrosine phosphatase (PTPase) activity of that found in HL-60 cells. This enzyme has a Km of 0.932 mmol/L and a Vmax of 0.355 mumol inorganic phosphate released/mg protein/min, which is similar to other cellular PTPase. Activation of PMN with f-Met-Leu-Phe and phorbol esters causes a slight but statistically significant drop in PMN PTPase activity. These results suggest that terminally differentiated myeloid cells have high tyrosine kinase and phosphatase activity, which may play a role in stimulus response coupling in the mature PMN.
D002460:Cell Line; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000201:enzymology* / Q000473:pathology; D009504:Neutrophils / Q000201:enzymology*; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D017027:Protein Tyrosine Phosphatases; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*
D011505:Protein-Tyrosine Kinases; D010749:Phosphoprotein Phosphatases; D017027:Protein Tyrosine Phosphatases