scholarly journals Protein tyrosine phosphorylation in rabbit peritoneal neutrophils

1990 ◽  
Vol 269 (2) ◽  
pp. 431-436 ◽  
Author(s):  
C K Huang ◽  
V Bonak ◽  
G R Laramee ◽  
J E Casnellie
Keyword(s):  
Tyrosine Phosphorylation ◽  
Kinase Inhibitor ◽  
Leukotriene B4 ◽  
Early Event ◽  
Ionophore A23187 ◽  
Protein Tyrosine Phosphorylation ◽  
Protein Tyrosine ◽  
Group A ◽  
Group B ◽  
Stimulation Of

Protein tyrosine phosphorylation in rabbit peritoneal neutrophils was examined by immunoblotting with antibodies specific for phosphotyrosine. Stimulation of the neutrophils with chemotactic factor fMet-Leu-Phe (10 nM) caused rapid increases of tyrosine phosphorylation of several proteins with apparent molecular masses of (Group A) 54-58 kDa and 100-125 kDa and (Group B) 36-41 kDa. Stimulation of Group A proteins was observed by fMet-Leu-Phe (10 nM, maximum at 20 s) and A23187 (1 microM, 1 min). Stimulation of Group B proteins was observed by fMet-Leu-Phe (ED50 0.15 nM, 1 min), leukotriene B4 (ED50 0.15 nM, 1 min), phorbol 12-myristate 13-acetate (PMA) (ED50 25 ng/ml, 10 min) and partially by ionophore A23187 (1 microM, 1 min). Pretreatment of the cell with the protein kinase inhibitor H-7 (25 microM, 5 min) and PMA (0.1 microgram/ml, 3 min) partially inhibited the fMet-Leu-Phe effect. However, pretreatment of the cells with quin 2/AM (20 microM, 10 min) completely inhibited the fMet-Leu-Phe effect. The results indicate that rapid regulation of tyrosine phosphorylation is an early event occurring in stimulated neutrophils. Furthermore the effect of fMet-Leu-Phe on tyrosine phosphorylation may require Ca2+ mobilization and may partially require the activity of H-7-sensitive protein kinases.

scholarly journals Tyrosine-specific protein phosphorylation during activation of human neutrophils

Blood ◽  
1990 ◽  
Vol 75 (12) ◽  
pp. 2445-2452 ◽  
Author(s):  
RL Berkow ◽  
RW Dodson
Keyword(s):  
Tyrosine Phosphorylation ◽  
Calcium Ionophore ◽  
Specific Protein ◽  
Leukotriene B4 ◽  
Calcium Ionophore A23187 ◽  
Human Neutrophils ◽  
Early Event ◽  
Ionophore A23187 ◽  
Protein Tyrosine Phosphorylation ◽  
Protein Tyrosine

Abstract The activation of human neutrophils by a variety of receptor-dependent and receptor-independent agonists induces the phosphorylation of a large number of proteins. Since we have previously shown that human neutrophils have at least two distinct tyrosine kinase activities, we examined protein tyrosine phosphorylation in human neutrophils stimulated with a variety of agonists. Using a monoclonal antibody specific for phosphotyrosine, the present study shows that the chemotactic peptides FMLP and leukotriene B4, the phorbol ester phorbol myristate acetate (PMA), and the calcium ionophore A23187 induce an increase in tyrosine phosphorylation of a number of neutrophil proteins. This increased protein tyrosine phosphorylation was dependent on the concentration of the agonist, as well as on the time of exposure to the agonist. Fractionation experiments showed that both a 150,000 g cytosolic and a particulate preparation showed increases in protein tyrosine phosphorylation with stimulation by FMLP or PMA, and showed that the pattern of protein tyrosine phosphorylation was slightly different in the FMLP- and PMA-stimulated cells. These data indicate that protein tyrosine phosphorylation is an early event in the activation of human neutrophils by a variety of receptor-dependent and receptor-independent agonists.

scholarly journals Tyrosine-specific protein phosphorylation during activation of human neutrophils

Blood ◽  
1990 ◽  
Vol 75 (12) ◽  
pp. 2445-2452
Author(s):  
RL Berkow ◽  
RW Dodson
Keyword(s):  
Tyrosine Phosphorylation ◽  
Calcium Ionophore ◽  
Specific Protein ◽  
Leukotriene B4 ◽  
Calcium Ionophore A23187 ◽  
Human Neutrophils ◽  
Early Event ◽  
Ionophore A23187 ◽  
Protein Tyrosine Phosphorylation ◽  
Protein Tyrosine

The activation of human neutrophils by a variety of receptor-dependent and receptor-independent agonists induces the phosphorylation of a large number of proteins. Since we have previously shown that human neutrophils have at least two distinct tyrosine kinase activities, we examined protein tyrosine phosphorylation in human neutrophils stimulated with a variety of agonists. Using a monoclonal antibody specific for phosphotyrosine, the present study shows that the chemotactic peptides FMLP and leukotriene B4, the phorbol ester phorbol myristate acetate (PMA), and the calcium ionophore A23187 induce an increase in tyrosine phosphorylation of a number of neutrophil proteins. This increased protein tyrosine phosphorylation was dependent on the concentration of the agonist, as well as on the time of exposure to the agonist. Fractionation experiments showed that both a 150,000 g cytosolic and a particulate preparation showed increases in protein tyrosine phosphorylation with stimulation by FMLP or PMA, and showed that the pattern of protein tyrosine phosphorylation was slightly different in the FMLP- and PMA-stimulated cells. These data indicate that protein tyrosine phosphorylation is an early event in the activation of human neutrophils by a variety of receptor-dependent and receptor-independent agonists.

Stimulation of protein tyrosine phosphorylation by platelet-activating factor and progesterone in human spermatozoa

1995 ◽  
Vol 108 (1-2) ◽  
pp. 35-42 ◽  
Author(s):  
Michaela Luconi ◽  
Lorella Bonaccorsi ◽  
Csilla Krausz ◽  
Ginetta Gervasi ◽  
Gianni Forti ◽  
...  
Keyword(s):  
Tyrosine Phosphorylation ◽  
Platelet Activating Factor ◽  
Human Spermatozoa ◽  
Protein Tyrosine Phosphorylation ◽  
Protein Tyrosine ◽  
Stimulation Of

scholarly journals Constitutive secretion of serum albumin requires reversible protein tyrosine phosphorylation events in trans-Golgi

2005 ◽  
Vol 289 (3) ◽  
pp. C748-C756 ◽  
Author(s):  
Rachel J. Webb ◽  
Jacob D. Judah ◽  
Lee-Chiang Lo ◽  
Geraint M. H. Thomas
Keyword(s):  
Serum Albumin ◽  
Tyrosine Phosphorylation ◽  
Tyrosine Kinases ◽  
Kinase Inhibitor ◽  
Tyrosine Phosphatase ◽  
Protein Tyrosine Phosphatases ◽  
Protein Tyrosine Phosphorylation ◽  
Albumin Secretion ◽  
Protein Tyrosine ◽  
Constitutive Secretion

Serum albumin secretion from rat hepatocytes proceeds via the constitutive pathway. Although much is known about the role of protein tyrosine phosphorylation in regulated secretion, nothing is known about its function in the constitutive process. Here we show that albumin secretion is inhibited by the tyrosine kinase inhibitor genistein but relatively insensitive to subtype-selective inhibitors or treatments. Secretion is also blocked in a physiologically identical manner by the tyrosine phosphatase inhibitors pervanadate and bisperoxo(1,10-phenanthroline)-oxovanadate. Inhibition of either the kinase(s) or phosphatase(s) leads to the accumulation of albumin between the trans-Golgi and the plasma membrane, whereas the immediate precursor proalbumin builds up in a proximal compartment. The trans-Golgi marker TGN38 is rapidly dispersed under conditions that inhibit tyrosine phosphatase action, whereas the distribution of the cis-Golgi marker GM130 is insensitive to genistein or pervanadate. By using a specifically reactive biotinylation probe, we detected protein tyrosine phosphatases in highly purified rat liver Golgi membranes. These membranes also contain both endogenous tyrosine kinases and their substrates, indicating that enzymes and substrates for reversible tyrosine phosphorylation are normal membrane-resident components of this trafficking compartment. In the absence of perturbation of actin filaments and microtubules, we conclude that reversible protein tyrosine phosphorylation in the trans-Golgi network is essential for albumin secretion and propose that the constitutive secretion of albumin is in fact a regulated process.

Delayed neuronal death in ischemic hippocampus involves stimulation of protein tyrosine phosphorylation

1996 ◽  
Vol 271 (4) ◽  
pp. C1085-C1097 ◽  
Author(s):  
T. Ohtsuki ◽  
M. Matsumoto ◽  
K. Kitagawa ◽  
T. Mabuchi ◽  
K. Mandai ◽  
...  
Keyword(s):  
Tyrosine Phosphorylation ◽  
Kainic Acid ◽  
Neuronal Death ◽  
Tyrosine Kinases ◽  
Ischemia Reperfusion ◽  
Delayed Neuronal Death ◽  
Mongolian Gerbils ◽  
Protein Tyrosine Phosphorylation ◽  
Protein Tyrosine ◽  
Stimulation Of

Glutamate triggers neuronal degeneration after ischemia-reperfusion in the brain. However, the details of intracellular signal transduction that propagates cell death remain unknown. The present work investigated whether protein tyrosine phosphorylation mediates neuronal death in the ischemic brain. Transient forebrain ischemia for 5-10 min in Mongolian gerbils or intoxication with the glutamate analogue kainic acid (12 mg/kg) in Sprague-Dawley rats caused neuronal death selectively in the hippocampus 2-4 days or 1 day later, respectively. Under these conditions, 160-, 115-, 105-, 92-, and 85-kDa proteins showed a significant increase in tyrosyl residue phosphorylation selectively in the hippocampus 3-12 h after ischemia or 4-8 h after kainic acid-induced seizures. Tyrosine kinases, including pp60c-src, were activated without a change of tyrosine phosphatases. Administration of radicicol, a selective inhibitor of tyrosine kinases, attenuated stimulation of tyrosine phosphorylation and hippocampal degeneration after ischemia or kainic acid injection. The results suggest that protein tyrosine phosphorylation might propagate delayed neuronal death in the mature hippocampus through glutamate overload after ischemia-reperfusion.

scholarly journals Tyrosine Phosphorylation and p72syk Activation by an Anti-Glycoprotein Ib Monoclonal Antibody

Blood ◽  
1997 ◽  
Vol 89 (5) ◽  
pp. 1590-1598 ◽  
Author(s):  
Mutsumasa Yanabu ◽  
Yukio Ozaki ◽  
Shosaku Nomura ◽  
Tetsuya Miyake ◽  
Yasuhiko Miyazaki ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Tyrosine Kinase ◽  
Tyrosine Phosphorylation ◽  
Kinase Inhibitor ◽  
Platelet Rich Plasma ◽  
Platelet Activating Factor ◽  
Protein Tyrosine Phosphorylation ◽  
Fab Fragment ◽  
Glycoprotein Ib ◽  
Protein Tyrosine

Abstract NNKY5-5, an IgG monoclonal antibody directed against the von Willebrand factor-binding domain of glycoprotein (GP) Ibα, induced weak but irreversible aggregation (or association) of platelets in citrate-anticoagulated platelet-rich plasma. This phenomenon was defined as small aggregate formation (SAF ). Platelets in hirudin-anticoagulated plasma or washed platelets showed little response to NNKY5-5 alone, but the antibody potentiated aggregation induced by low concentrations of adenosine diphosphate or platelet-activating factor. NNKY5-5 did not induce granule release or intracellular Ca2+ mobilization. However, NNKY5-5 caused tyrosine phosphorylation of a 64-kD protein and activation of a tyrosine kinase, p72syk. An anti-FcγII receptor antibody had no effect on SAF, suggesting that NNKY5-5 activated platelets by interacting with glycoprotein Ib. Fab′ fragments of NNKY5-5 did not induce SAF, but potentiated aggregation induced by other agonists. The Fab′ fragment of NNKY5-5 induced the activation of p72syk, suggesting that such activation was independent of the FcγII receptor. Cross-linking of the receptor-bound Fab′ fragment of NNKY5-5 with a secondary antibody induced SAF. GRGDS peptide, chelation of extracellular Ca2+, and an anti-GPIIb/IIIa antibody inhibited NNKY5-5-induced SAF, but had no effect on 64-kD protein tyrosine phosphorylation or p72syk activations. Various inhibitors, including aspirin and protein kinase C, had no effect on SAF, protein tyrosine phosphorylation, or p72syk activation. In contrast, tyrphostin 47, a potent tyrosine kinase inhibitor, inhibited NNKY5-5–induced SAF as well as tyrosine phosphorylation and p72syk activation. Our findings suggest that binding of NNKY5-5 to GPIb potentiates platelet aggregation by facilitating the interaction between fibrinogen and GPIIb/IIIa through a mechanism associated with p72syk activation and tyrosine phosphorylation of a 64-kD protein.

Bradykinin stimulates the ERK→Elk-1→Fos/AP-1 pathway in mesangial cells

1998 ◽  
Vol 275 (3) ◽  
pp. F343-F352 ◽  
Author(s):  
Samir S. El-Dahr ◽  
Susana Dipp ◽  
William H. Baricos
Keyword(s):  
Transcription Factor ◽  
Tyrosine Kinase ◽  
Dna Synthesis ◽  
Tyrosine Phosphorylation ◽  
Kinase Inhibitor ◽  
Mesangial Cells ◽  
Fold Increase ◽  
Antisense Oligodeoxynucleotide ◽  
Protein Tyrosine Phosphorylation ◽  
Protein Tyrosine

Among its diverse biological actions, the vasoactive peptide bradykinin (BK) induces the transcription factor AP-1 and proliferation of mesangial cells (S. S. El-Dahr, S. Dipp, I. V. Yosipiv, and W. H. Baricos. Kidney Int. 50: 1850–1855, 1996). In the present study, we examined the role of protein tyrosine phosphorylation and the mitogen-activated protein kinases, ERK1/2,in mediating BK-induced AP-1 and DNA replication in cultured rat mesangial cells. BK (10−9 to 10−7 M) stimulated a rapid increase in tyrosine phosphorylation of multiple proteins with an estimated molecular mass of 120–130, 90–95, and 44–42 kDa. Immunoblots using antibodies specific for ERK or tyrosine-phosphorylated ERK revealed a shifting of p42 ERK2 to a higher molecular weight that correlated temporally with an increase in tyrosine-phosphorylated ERK2. Genistein, a specific tyrosine kinase inhibitor, prevented the phosphorylation of ERK2 by BK. In-gel kinase assays indicated that BK-induced tyrosine phosphorylation of ERK2 is accompanied by fourfold activation of its phosphotransferase activity toward the substrate PHAS-I ( P < 0.05). Furthermore, BK stimulated a 2.5-fold increase ( P < 0.05) in phosphorylation of Elk-1, a transcription factor required for growth factor-induced c-fos transcription. In accord with the stimulation of Elk-1 phosphorylation, BK induced c-fos gene expression and the production of Fos/AP-1 complexes. In addition, thymidine incorporation into DNA increased twofold ( P < 0.05) following BK stimulation. Each of these effects was blocked by tyrosine kinase inhibition with genistein or herbimycin A. Similarly, antisense oligodeoxynucleotide targeting of ERK1/2 mRNA inhibited BK-stimulated DNA synthesis. In contrast, protein kinase C inhibition or depletion had no effect on BK-induced c-fos mRNA, AP-1-DNA binding activity, or DNA synthesis. Collectively, these data demonstrate that BK activates the ERK→Elk-1→AP-1 pathway and that BK mitogenic signaling is critically dependent on protein tyrosine phosphorylation.

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