scholarly journals Identification of a Membrane Ig-Induced p38 Mitogen-Activated Protein Kinase Module That Regulates cAMP Response Element Binding Protein Phosphorylation and Transcriptional Activation in CH31 B Cell Lymphomas

2000 ◽  
Vol 164 (5) ◽  
pp. 2311-2319 ◽  
Author(s):  
Jennifer M. Swart ◽  
Daniele M. Bergeron ◽  
Thomas C. Chiles
Keyword(s):  
Protein Kinase ◽  
B Cell ◽  
Protein Phosphorylation ◽  
Transcriptional Activation ◽  
Camp Response Element Binding ◽  
Mitogen Activated Protein Kinase ◽  
Camp Response Element ◽  
Response Element ◽  
Mitogen Activated Protein ◽  
Element Binding Protein

scholarly journals A new phosphospecific cell-based ELISA for p42/p44 mitogen-activated protein kinase (MAPK), p38 MAPK, protein kinase B and cAMP-response-element-binding protein

2000 ◽  
Vol 350 (3) ◽  
pp. 717-722 ◽  
Author(s):  
Henri H. VERSTEEG ◽  
Evert NIJHUIS ◽  
Gijs R. VAN DEN BRINK ◽  
Maaike EVERTZEN ◽  
Gwenda N. PYNAERT ◽  
...  
Keyword(s):  
Signal Transduction ◽  
Protein Kinase ◽  
P38 Mapk ◽  
Protein Kinase B ◽  
Camp Response Element Binding ◽  
Mitogen Activated Protein Kinase ◽  
Camp Response Element ◽  
Response Element ◽  
Mitogen Activated Protein ◽  
Element Binding Protein

Assaying activation of signal transduction is laborious and does not allow the study of large numbers of samples, essential for high-throughput drug screens or for large groups of patients. Using phosphospecific antibodies, we have developed ELISA techniques enabling non-radioactive semi-quantitative assessment of the activation state of p42/p44 mitogen-activated protein kinase (MAPK), p38 MAPK, protein kinase B and the transcription factor cAMP-response-element-binding protein (CREB) in 96-well plates. This assay has been termed PACE (phosphospecific antibody cell-based ELISA) and was used successfully for both adherent and suspension cells. Various stimuli induced dose-dependent enzymic activity of which the kinetics closely correlated with those measured via classical methodology. Using PACE we have now characterized for the first time the concentration-dependent effects of various inflammatory prostaglandins on CREB phosphorylation in macrophages. PACE is a straightforward and novel technique enabling the large-scale analysis of signal transduction.

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