Stimulation of a Histone H4 Protein Kinase in Triton X-100 Lysates of Rabbit Peritoneal Neutrophils Pretreated With Chemotactic Factors: Effect of Leukotriene B4 and Cytochalasin B

1991 ◽  
Vol 49 (2) ◽  
pp. 158-162 ◽  
Author(s):  
Chi-Kuang Huang ◽  
Gary R. Laramee
Keyword(s):  
Protein Kinase ◽  
Cytochalasin B ◽  
Leukotriene B4 ◽  
Histone H4 ◽  
Chemotactic Factors ◽  
Triton X ◽  
Stimulation Of

scholarly journals Effects of chemotactic factors and other agents on the amounts of actin and a 65,000-mol-wt protein associated with the cytoskeleton of rabbit and human neutrophils.

1985 ◽  
Vol 101 (1) ◽  
pp. 182-188 ◽  
Author(s):  
R Yassin ◽  
J Shefcyk ◽  
J R White ◽  
W Tao ◽  
M Volpi ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Leukotriene B4 ◽  
Intracellular Concentration ◽  
Human Neutrophils ◽  
Free Calcium ◽  
High Osmolarity ◽  
Kinase C ◽  
Chemotactic Factors ◽  
Stimulation Of

Stimulation of rabbit neutrophils by the chemotactic factors fMet-Leu-Phe and leukotriene B4, by platelet activating factor, or by arachidonic acid produces a rapid and dose-dependent increase in the amounts of actin and of a 65,000-mol-wt protein associated with the cytoskeleton. Phorbol 12-myristate, 13-acetate, the calcium ionophore A23187 in the presence or absence of EGTA, and the fluorescent calcium chelator quin-2 also cause an increase in cytoskeletal actin. The stimulated increases in the cytoskeletal actin are not dependent on a rise in the intracellular concentration of free calcium and are not mediated by an increase in the intracellular pH or activation of protein kinase C. The increases in the cytoskeletal actin produced by fMet-Leu-Phe and leukotriene B4, but not by phorbol 12-myristate, 13-acetate, are inhibited by high osmolarity. The effect of hyperosmolarity requires a decrease in cell volume, is not mediated by an increase in basal intracellular concentration of free calcium, and is not prevented by pretreating the cells with amiloride. Preincubation of the cells with hyperosmotic solution also inhibits degranulation produced by all the stimuli tested. The inhibitory action of high osmolarity on the fMet-Leu-Phe and leukotriene B4 induced stimulation of cytoskeletal actin is discussed in terms of the possibility that the addition of high osmolarity, either directly or through activation of protein kinase C, causes receptor uncoupling.

scholarly journals Protein kinase regulation of a cloned epithelial Na+ channel.

1996 ◽  
Vol 108 (1) ◽  
pp. 49-65 ◽  
Author(s):  
M S Awayda ◽  
I I Ismailov ◽  
B K Berdiev ◽  
C M Fuller ◽  
D J Benos
Keyword(s):  
Protein Kinase ◽  
Lipid Bilayers ◽  
Xenopus Oocytes ◽  
Cytochalasin B ◽  
Na Channel ◽  
Planar Lipid Bilayers ◽  
Open Probability ◽  
Alpha Beta ◽  
Epithelial Na Channel ◽  
Stimulation Of

We examined the regulation of a cloned epithelial Na+ channel (alpha beta gamma-rENaC) by protein kinase A (PKA) and protein kinase C (PKC). Experiments were performed in Xenopus oocytes and in planar lipid bilayers. At a holding potential of -100 mV, amiloride-sensitive current averaged -1,279 +/- 111 nA (n = 7) in alpha beta gamma-rENaC-expressing oocytes. Currents in water-injected oocytes were essentially unresponsive to 10 microM amiloride. A 1-h stimulation of PKC with 100 nM of PMA inhibited whole-cell currents in Xenopus oocytes to 17.1 +/- 1.8, and 22.1 +/- 2.6% of control (n = 7), at holding potentials of -100 and +40 mV, respectively. Direct injection of purified PKC resulted in similar inhibition to that observed with PMA. Additionally, the inactive phorbol ester, phorbol-12-myristate-13-acetate, 4-O-methyl, was without effect on alpha beta gamma-rENaC currents. Pretreatment with the microtubule inhibitor colchicine (100 microM) did not modify the inhibitory effect of PMA; however, pretreatment with 20 microM cytochalasin B decreased the inhibitory action of PMA to < 20% of that previously observed. In vitro-synthesized alpha beta gamma-rENaC formed an amiloride-sensitive Na(+)-selective channel when incorporated into planar lipid bilayers. Addition of PKC, diacyl-glycerol, and Mg-ATP to the side opposite that which amiloride blocked, decreased the channel's open probability (Po) from 0.44 +/- 0.06 to 0.13 +/- 0.03 (n = 9). To study the effects of PKA on alpha beta gamma-rENaC expressed in Xenopus oocytes, cAMP levels were elevated with 10 microM forskolin and 1 mM isobutyl-methyl-xanthine. This cAMP-elevating cocktail did not cause any stimulation of alpha beta gamma-rENaC currents in either the inward or outward directions. This lack of activation was also observed in oocytes preinhibited with PMA and in oocytes pretreated with cytochalasin B and PMA. Neither alpha-rENaC nor alpha beta gamma-rENaC incorporated into planar lipid bilayers could be activated with PKA and Mg-ATP added to either side of the membrane, as Po remained at 0.63 +/- 0.06 (n = 7) and 0.45 +/- 0.05 (n = 9), respectively. We conclude that: alpha beta gamma-rENaC is inhibited by PKC, and that alpha beta gamma-rENaC is not activated by PKA.

Diacylglycerol kinase inhibitor R59022 and stimulated neutrophil responses

1988 ◽  
Vol 255 (5) ◽  
pp. C589-C594 ◽  
Author(s):  
J. L. Mege ◽  
W. Tao ◽  
T. F. Molski ◽  
J. Gomez-Cambronero ◽  
C. K. Huang ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Cytochalasin B ◽  
Kinase Inhibitor ◽  
Platelet Activating Factor ◽  
Diacylglycerol Kinase ◽  
Leukotriene B4 ◽  
Free Calcium ◽  
Kinase C ◽  
Protein Kinase C Inhibitor

The generation of phosphatidic acid in neutrophils stimulated by the chemotactic factor formylmethionyl-leucyl-phenylalanine (fMet-Leu-Phe) is inhibited by the diacylglycerol kinase inhibitor R59022. Superoxide generation produced by fMet-Leu-Phe, leukotriene B4, platelet-activating factor, or phorbol 12-myristate 13-acetate can be greatly increased in neutrophils pretreated with R59022. The potentiation occurs in the presence or absence of cytochalasin B and is evident in the absence of extracellular calcium. In addition, where the superoxide generated by fMet-Leu-Phe is not inhibited by the protein kinase C inhibitor 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7), the increase by R59022 is diminished by this compound. Unlike cytochalasin B, R59022 does not affect the increase in cytoskeletal actin produced by fMet-Leu-Phe or platelet-activating factor nor does it decrease the basal level. Furthermore, the basal intracellular concentration of free calcium, but not the rise produced by fMet-Leu-Phe or platelet-activating factor, is elevated by R59022. The data presented here suggest that the potentiation by R59022 of the oxidative burst is most likely mediated through protein kinase C.

scholarly journals Protein kinase activity in membrane preparations from ox brain. Stimulation of intrinsic activity by adenosine 3′:5′-cyclic monophosphate

1973 ◽  
Vol 132 (3) ◽  
pp. 483-492 ◽  
Author(s):  
Malcolm Weller ◽  
Richard Rodnight
Keyword(s):  
Protein Kinase ◽  
Cyclic Amp ◽  
Time Course ◽  
Kinase Activity ◽  
Intrinsic Activity ◽  
Protein Kinase Activity ◽  
Ph Optimum ◽  
Basal Activity ◽  
Triton X ◽  
Stimulation Of

1. Properties of the stimulation by cyclic AMP of the intrinsic protein kinase activity of membrane fragments from ox brain were studied. 2. Stimulation of activity declined from about 100% at 1min to less than 20% at 10min. The time-course was explained by the observation that cyclic AMP did not stimulate turnover of protein-bound serine phosphate once the membrane protein was fully phosphorylated. 3. Cyclic AMP accelerated the activity of a component of the basal activity rather than activating a different kinase. 4. The pH optimum for both the stimulated and basal activities was 7.2–7.4. NaCl (100mm) and KCl (10–100mm) inhibited the stimulated activity but did not affect the basal activity. 5. Strychnine and theophylline inhibited both activites equally, but the stimulated activity was more sensitive to inhibition by adenosine, bicuculline, vinblastin, veratrine, N-ethylmaleimide and cysteine. 6. No firm evidence for a role for endogenous cyclic AMP in the basal activity was found, but the possibility was not excluded. 7. Some 90% of both the stimulated and basal activities remained in an insoluble form after treatment of the membrane fragments with Triton X-100 (0.5%).

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