Generation of an active protein-tyrosine kinase from lymphocytes by proteolysis.
The major NaCl-stimulated protein-tyrosine kinase activity found in soluble thymus extracts, as measured by the phosphorylation of angiotensin I, is a 40-kDa enzyme known as p40 (Zioncheck, T. F., Harrison, M. L., and Geahlen, R. L. (1986) J. Biol. Chem. 261, 15637-15643). Antibodies prepared against p40 cross-react with a 72-kDa protein-tyrosine kinase (p72) from spleen or thymus that is closely related to p40 by peptide-mapping experiments. The recovery of p40 from spleen homogenates is reduced while the recovery of p72 is enhanced by the addition of high concentrations of leupeptin or soybean trypsin inhibitor to the homogenization media. The generation of p40 in spleen homogenates occurs with a concomitant increase in protein-tyrosine kinase activity. Activated catalytic fragments of 38-43 kDa can be generated by the treatment of partially purified p72 with trypsin or papain. The p72 protein-tyrosine kinase is found at the highest levels in spleen, thymus, and lung, tissues that also have high protein-tyrosine kinase activity and generate high levels of p40 following homogenization. p72 is also found in certain T and B cell-derived cell lines and in NIH3T3 cells.
Zioncheck
T F
TF
Department of Medicinal Chemistry and Pharmacognosy, School of Pharmacy and Pharmacal Sciences, Purdue University, West Lafayette, Indiana 47907.
Harrison
M L
ML
Isaacson
C C
CC
Geahlen
R L
RL
D000803:Angiotensin I / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D007527:Isoenzymes / Q000097:blood; D008214:Lymphocytes / Q000201:enzymology*; D008970:Molecular Weight; D010449:Peptide Mapping; D010766:Phosphorylation; D011480:Protease Inhibitors / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000097:blood*; D012965:Sodium Chloride / Q000494:pharmacology
D007527:Isoenzymes; D011480:Protease Inhibitors; D012965:Sodium Chloride; D000803:Angiotensin I; D011505:Protein-Tyrosine Kinases