Partial characterization of protein tyrosine kinases in human lymphoid cells.
We have examined several aspects of protein tyrosine kinase (PTK)-activity in the cytosolic and membrane fractions of both normal and malignant lymphoid cells. The expression of the PTK-encoding oncogenes fes, abl and src, was investigated with the use of antibodies generated to their respective gene products. These antibodies recognized proteins in all cell types examined, most frequently in both the cytosolic and the membrane fraction. PTKs were partially characterized by FPLC. PTK-activities of column fractions were assayed using a non-radioactive dot blot assay. Cytosolic and membrane fractions showed FPLC patterns with a constant as well as a variable part in both normal and malignant cells, suggestive of PTKs with specialized functions in normal cell growth and transformation. Lastly, using antibodies to phosphotyrosine, we found that cytosolic fractions contained the majority of proteins phosphorylated at tyrosine in all cell types. Normal peripheral blood lymphocytes and B-lymphoma cells showed a great similarity in tyrosine phosphorylation pattern, while in tonsillar lymphocytes a clearly different pattern was found. These methods further characterize PTK-activities in lymphoid cells, and the results give evidence that PTKs in normal and malignant cells have both similar and different aspects.
Punt
C J
CJ
Department of Medical Oncology, University Hospital Nijmegen, The Netherlands.
Rijksen
G
G
van Oirschot
B A
BA
Kester-Schipper
G P
GP
van der Heijden
M C
MC
Dekker
A W
AW
Staal
G E
GE
D002462:Cell Membrane / Q000201:enzymology; D003600:Cytosol / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008214:Lymphocytes / Q000276:immunology*; D016393:Lymphoma, B-Cell / Q000201:enzymology; D008970:Molecular Weight; D009856:Oncogene Proteins, Viral / Q000032:analysis*; D009857:Oncogenes; D014066:Palatine Tonsil / Q000276:immunology; D010750:Phosphoproteins / Q000302:isolation & purification; D019000:Phosphotyrosine; D011505:Protein-Tyrosine Kinases / Q000032:analysis* / Q000097:blood / Q000302:isolation & purification; D016315:Proto-Oncogene Proteins c-abl / Q000032:analysis*; D014443:Tyrosine / Q000031:analogs & derivatives / Q000032:analysis
D009856:Oncogene Proteins, Viral; D010750:Phosphoproteins; D019000:Phosphotyrosine; D014443:Tyrosine; D011505:Protein-Tyrosine Kinases; D016315:Proto-Oncogene Proteins c-abl