scholarly journals Stimulation of tyrosine phosphorylation and mitogen-activated-protein (MAP) kinase activity in human SH-SY5Y neuroblastoma cells by carbachol

1993 ◽  
Vol 294 (2) ◽  
pp. 545-550 ◽  
Author(s):  
S Offermanns ◽  
E Bombien ◽  
G Schultz
Keyword(s):  
Map Kinase ◽  
Tyrosine Phosphorylation ◽  
Kinase Activity ◽  
Neuroblastoma Cells ◽  
Anion Exchange Column ◽  
Human Neuroblastoma ◽  
Protein Map ◽  
Mitogen Activated Protein ◽  
M3 Receptor ◽  
Stimulation Of

Activation of the G-protein-coupled muscarinic (M3) receptor in human neuroblastoma SH-SY5Y cells is known to lead to phosphoinositol hydrolysis and noradrenaline release. In this study, the effect of carbachol on tyrosine phosphorylation and mitogen-activated protein (MAP) kinase activity in SH-SY5Y cells was examined. Carbachol concentration-dependently induced tyrosine phosphorylation of several proteins, including one of 42 kDa. This tyrosine-phosphorylated 42 kDa protein co-eluted from a Mono Q anion-exchange column with MAP kinase activity and with immunologically detected MAP kinase. Stimulation of tyrosine phosphorylation and activation of MAP kinase were also observed after incubation of cells with phorbol 12-myristate 13-acetate (PMA) and epidermal growth factor (EGF). Down-regulation or inhibition of protein kinase C (PKC) abolished the stimulatory effects of both carbachol and PMA on MAP kinase activity, whereas EGF-stimulated MAP kinase activity remained unaffected. Thus carbachol acting through the muscarinic (M3) receptor PKC-dependently induced tyrosine phosphorylation and activation of a 42 kDa MAP kinase in SH-SY5Y cells, whereas EGF-induced MAP kinase activation occurred independently of PKC.

Biochemical characterization of mitogen-activated protein (MAP) kinase activity in Toxoplasma gondii

2000 ◽  
Vol 86 (7) ◽  
pp. 588-598 ◽  
Author(s):  
Marie-Paule Roisin ◽  
Florence Robert-Gangneux ◽  
Claudine Creuzet ◽  
Jean Dupouy-Camet
Keyword(s):  
Toxoplasma Gondii ◽  
Map Kinase ◽  
Kinase Activity ◽  
Biochemical Characterization ◽  
Protein Map ◽  
Mitogen Activated Protein

scholarly journals Selective activation of p42 mitogen-activated protein (MAP) kinase in murine B lymphoma cell lines by membrane immunoglobulin cross-linking. Evidence for protein kinase C-independent and -dependent mechanisms of activation

1992 ◽  
Vol 287 (1) ◽  
pp. 269-276 ◽  
Author(s):  
M R Gold ◽  
J S Sanghera ◽  
J Stewart ◽  
S L Pelech
Keyword(s):  
Map Kinase ◽  
Tyrosine Phosphorylation ◽  
Cell Lines ◽  
Map Kinases ◽  
Phorbol Esters ◽  
Cross Linking ◽  
Kinase Activation ◽  
Membrane Immunoglobulin ◽  
Protein Map ◽  
Mitogen Activated Protein

Cross-linking of membrane immunoglobulin (mIg), the B lymphocyte antigen receptor, with anti-receptor antibodies stimulates tyrosine phosphorylation of a number of proteins, including one of 42 kDa. Proteins with a similar molecular mass are tyrosine-phosphorylated in response to receptor stimulation in other cell types and have been identified as serine/threonine kinases, termed mitogen-activated protein (MAP) kinases or extracellular signal-regulated kinases (ERKs). The MAP kinases constitute a family of related kinases, at least three of which have molecular masses of 40-45 kDa. In this paper we show that mIg cross-linking stimulated the myelin basic protein phosphotransferase activity characteristic of MAP kinase in both mature and immature murine B cell lines. This enzyme activity co-purified on three different columns with a 42 kDa protein that was tyrosine-phosphorylated (pp42) in response to mIg cross-linking and which reacted with a panel of anti-(MAP kinase) antibodies. Although immunoblotting with the anti-(MAP kinase) antibodies showed that these B cell lines expressed both 42 kDa and 44 kDa forms of MAP kinase, only the 42 kDa form was activated and tyrosine-phosphorylated to a significant extent. Activation of protein kinase C (PKC) with phorbol esters also resulted in selective tyrosine phosphorylation and activation of the 42 kDa MAP kinase. This suggested that mIg-induced MAP kinase activation could be due to stimulation of PKC by mIg. However, mIg-stimulated MAP kinase activation and pp42 tyrosine phosphorylation was only partially blocked by a PKC inhibitor, the staurosporine analogue Compound 3. In contrast, Compound 3 completely blocked the ability of phorbol esters to stimulate MAP kinase activity and induce tyrosine phosphorylation of pp42. Thus mIg may activate MAP kinase by both PKC-dependent and -independent mechanisms.

MITOGEN-ACTIVATED PROTEIN (MAP) KINASE ACTIVITY INCREASES AFTER FOCAL CEREBRAL ISCHEMIA IN RATS

1998 ◽  
Vol 26 (Supplement) ◽  
pp. 84A
Author(s):  
John W. Kuluz ◽  
Charles L. Schleien ◽  
Y. Kang ◽  
Joseph Neary
Keyword(s):  
Cerebral Ischemia ◽  
Map Kinase ◽  
Kinase Activity ◽  
Focal Cerebral Ischemia ◽  
Protein Map ◽  
Mitogen Activated Protein

scholarly journals Neuroprotective Agent Chlomethiazole Attenuates c-fos, c-jun, and AP-1 Activation through Inhibition of p38 MAP Kinase

2000 ◽  
Vol 20 (7) ◽  
pp. 1077-1088 ◽  
Author(s):  
Anastasia Simi ◽  
Magnus Ingelman-Sundberg ◽  
Niclas Tindberg
Keyword(s):  
Cerebral Ischemia ◽  
Map Kinase ◽  
Neuroblastoma Cells ◽  
P38 Map Kinase ◽  
Luciferase Reporter ◽  
Human Neuroblastoma ◽  
Chloride Currents ◽  
Neuroprotective Agent ◽  
Luciferase Reporter Plasmid ◽  
Mitogen Activated Protein

Recent evidence suggests that stress-activated protein kinases expressed in glial cells have very important roles during cerebral ischemia. The neuroprotective agent chlomethiazole, which is known to enhance the conductance at the GABAA receptor complex, is presently in clinical trials for the treatment of severe stroke. Here the authors suggested that chlormethiazole has anti-inflammatory properties because it potently and selectively inhibited p38 mitogen-activated protein (MAP) kinase in primary cortical glial cultures. The inhibition of p38 MAP kinase resulted in the attenuation of the induction of c-fos and c-jun mRNA and AP-1 DNA binding by lipopolysaccharide (LPS). In addition, chlomethiazole inhibited the activation of an AP-1-dependent luciferase reporter plasmid in SK-N-MC human neuroblastoma cells in response to glutamate. Chlomethiazole inhibited the p38 MAP kinase activity as revealed by the decrease in the LPS-induced phosphorylation of the substrates ATF-2 and hsp27, whereas the phosphorylation status of the p38 MAP kinase itself was unaffected. Interestingly, chlomethiazole exhibited an IC50 of ~ 2 μmol/L for inhibition of c-fos mRNA expression, indicating 25 to 75 times higher potency than reported EC50 values for enhancing GAB AA chloride currents. The results indicated a novel mechanism of action of chlomethiazole, and provided support for a distinctive role of p38 MAP kinase in cerebral ischemia.

scholarly journals c-Jun N-terminal phosphorylation correlates with activation of the JNK subgroup but not the ERK subgroup of mitogen-activated protein kinases.

1994 ◽  
Vol 14 (10) ◽  
pp. 6683-6688 ◽  
Author(s):  
A Minden ◽  
A Lin ◽  
T Smeal ◽  
B Dérijard ◽  
M Cobb ◽  
...  
Keyword(s):  
Map Kinase ◽  
Map Kinases ◽  
New Members ◽  
Protein Map ◽  
Mitogen Activated Protein ◽  
Extracellular Stimuli ◽  
Terminal Site ◽  
Stimulation Of

c-Jun transcriptional activity is stimulated by phosphorylation at two N-terminal sites: Ser-63 and -73. Phosphorylation of these sites is enhanced in response to a variety of extracellular stimuli, including growth factors, cytokines, and UV irradiation. New members of the mitogen-activated protein (MAP) kinase group of signal-transducing enzymes, termed JNKs, bind to the activation domain of c-Jun and specifically phosphorylate these sites. However, the N-terminal sites of c-Jun were also suggested to be phosphorylated by two other MAP kinases, ERK1 and ERK2. Despite these reports, we find that unlike the JNKs, ERK1 and ERK2 do not phosphorylate the N-terminal sites of c-Jun in vitro; instead they phosphorylate an inhibitory C-terminal site. Furthermore, the phosphorylation of c-Jun in vivo at the N-terminal sites correlates with activation of the JNKs but not the ERKs. The ERKs are probably involved in the induction of c-fos expression and thereby contribute to the stimulation of AP-1 activity. Our study suggests that two different branches of the MAP kinase group are involved in the stimulation of AP-1 activity through two different mechanisms.

Lysophosphatidylcholine activates mesangial cell PKC and MAP kinase by PLCγ-1 and tyrosine kinase-Ras pathways

1999 ◽  
Vol 277 (3) ◽  
pp. F328-F337 ◽  
Author(s):  
Babu V. Bassa ◽  
Daeyoung D. Roh ◽  
Nosratola D. Vaziri ◽  
Michael A. Kirschenbaum ◽  
Vaijinath S. Kamanna
Keyword(s):  
Tyrosine Kinase ◽  
Map Kinase ◽  
Protein Tyrosine Kinase ◽  
Kinase Activity ◽  
Mesangial Cells ◽  
Kinase Activation ◽  
Kinase C ◽  
Protein Map ◽  
Mitogen Activated Protein ◽  
Pkc Activation

Although lysophosphatidylcholine (LPC)-mediated cellular responses are attributed to the activation of protein kinase C (PKC), relatively little is known about the upstream signaling mechanisms that regulate the activation of PKC and downstream mitogen-activated protein (MAP) kinase. LPC activated p42 MAP kinase and PKC in mesangial cells. LPC-mediated MAP kinase activation was inhibited (but not completely) by PKC inhibition, suggesting additional signaling events. LPC stimulated protein tyrosine kinase (PTK) activity and induced Ras-GTP binding. LPC-induced MAP kinase activity was blocked by the PTK inhibitor genistein. Because LPC increased PTK activity, we examined the involvement of phospholipase Cγ-1 (PLCγ-1) as a key participant in LPC-induced PKC activation. LPC stimulated the phosphorylation of PLCγ-1. PTK inhibitors suppressed LPC-induced PKC activity, whereas the same had no effect on phorbol 12-myristate 13-acetate-mediated PKC activity. Other lysophospholipids [e.g., lysophosphatidylinositol and lysophosphatidic acid (LPA)] also induced MAP kinase activity, and only LPA-induced MAP kinase activation was sensitive to pertussis toxin. These results indicate that LPC-mediated PKC activation may be regulated by PTK-dependent activation of PLCγ-1, and both PKC and PTK-Ras pathways are involved in LPC-mediated downstream MAP kinase activation.

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