Calcium-induced p56(Lck) phosphorylation in human T lymphocytes via calmodulin dependent kinase.
We report that stimulation of both primary human and Jurkat T lymphocytes with the calcium ionophore ionomycin, or A23187, results in the phosphorylation of p56(Lck) as determined by shifts in mobility of p56(Lck) on immunoblots. The shifts in the mobility of p56(Lck) induced by ionomycin could be blocked by preincubation of the cells with EGTA, demonstrating the requirement for extracellular calcium in this response. Although increases in intracellular calcium have been shown to modulate CD45 activity, phosphorylation of p56(Lck) was not mediated via CD45. Ionomycin stimulation of J45. 01 cells, a CD45-negative Jurkat cell derivative, also resulted in p56(Lck) mobility shifts. Instead, this response appears to be mediated via a calmodulin-dependent kinase. This response could be blocked by calmidazolium, an inhibitor of calmodulin, and KN-93, an inhibitor of calmodulin-dependent kinases (CaM-Kinase). KN-92, an inactive analog of KN-93, failed to block this response. These studies demonstrate a new role for calcium and CaM-Kinase in human T-lymphocytes and describe a novel mechanism by which p56(Lck) can be modulated.
Franklin
R A
RA
Department of Microbiology and Immunology, East Carolina University School of Medicine, Greenville, North Carolina, 27858, USA.
Mcleod
A
A
Robinson
P J
PJ
D001596:Benzylamines / Q000494:pharmacology; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D015759:Ionomycin / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D019169:Jurkat Cells; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / Q000378:metabolism*; D010766:Phosphorylation; D013449:Sulfonamides / Q000494:pharmacology; D013601:T-Lymphocytes / Q000201:enzymology / Q000378:metabolism*; D000096926:Benzenesulfonamides
D001596:Benzylamines; D000001:Calcimycin; D002118:Calcium; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D004791:Enzyme Inhibitors; D015759:Ionomycin; D007476:Ionophores; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D013449:Sulfonamides; C072105:KN 93; D000096926:Benzenesulfonamides