|
|
|
|||||||
|
|||||||||
J. Biol. Chem., Vol. 275, Issue 43, 33536-33541, October 27, 2000
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
From the Many signal transduction pathways are mediated by
the second messengers cGMP and cAMP, cGMP- and
cAMP-dependent protein kinases (cGK and PKA),
phosphodiesterases, and ion channels. To distinguish among the
different cGMP effectors, inhibitors of cGK and PKA have been developed
including the K-252 compound KT5823 and the isoquinolinesulfonamide
H89. KT5823, an in vitro inhibitor of cGK, has also been
used in numerous studies with intact cells to implicate or rule out the
involvement of this protein kinase in a given cellular response.
However, the efficacy and specificity of KT5823 as cGK inhibitor in
intact cells or tissues have never been demonstrated. Here, we analyzed
the effects of both KT5823 and H89 on cyclic-nucleotide-mediated
phosphorylation of vasodilator-stimulated phosphoprotein (VASP) in
intact human platelets and rat mesangial cells. These two cell types
both express high levels of cGK. KT5823 inhibited purified cGK.
However, with both intact human platelets and rat mesangial cells,
KT5823 failed to inhibit cGK-mediated serine 157 and serine 239 phosphorylation of VASP induced by nitric oxide, atrial natriuretic
peptide, or the membrane-permeant cGMP analog, 8-pCPT-cGMP. KT5823
enhanced 8-pCPT-cGMP-stimulated VASP phosphorylation in platelets and
did not inhibit forskolin-stimulated VASP phosphorylation in either
platelets or mesangial cells. In contrast H89, an inhibitor of both PKA
and cGK, clearly inhibited 8-pCPT-cGMP and forskolin-stimulated VASP
phosphorylation in the two cell types. The data indicate that KT5823
inhibits purified cGK but does not affect a cGK-mediated response in
the two different cell types expressing cGK I. These observations
indicate that data that interpret the effects of KT5823 in
intact cells as the major or only criteria supporting the involvement
of cGK clearly need to be reconsidered.
KT5823 Inhibits cGMP-dependent Protein Kinase
Activity in Vitro but Not in Intact Human Platelets and Rat
Mesangial Cells*
§,§¶,¶,,,,
,, and
Institute of Clinical Biochemistry and
Pathobiochemistry and the Division of Nephrology, Medical
University Clinic Wuerzburg, 97080 Wuerzburg, Germany, the
¶ Sechenov Institute of Evolutionary Physiology and Biochemistry,
Academy of Sciences, 194223 St. Petersburg, Russia, and the ** Division
of Molecular and Cellular Pathology, University of Alabama, Birmingham,
Alabama 35294-0019.
*
This work was supported by the Deutsche
Forschungsgemeinschaft (SFB 355) and by the Wilhelm Sander Stiftung.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Institut für
Klinische Biochemie und Pathobiochemie, Medizinische
Universitaetsklinik, Josef-Schneider Str. 2, D-97080 Wuerzburg,
Germany. Tel.: 49-931-201-2782; Fax: 49-931-201-3137; E-mail:
palme@klin-biochem.uni-wuerzburg.de.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  N. Valtcheva, P. Nestorov, A. Beck, M. Russwurm, M. Hillenbrand, P. Weinmeister, and R. Feil The Commonly Used cGMP-dependent Protein Kinase Type I (cGKI) Inhibitor Rp-8-Br-PET-cGMPS Can Activate cGKI in Vitro and in Intact Cells J. Biol. Chem., January 2, 2009; 284(1): 556 - 562. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X.-m. Liu, K. J. Peyton, N. N. Mendelev, H. Wang, D. A. Tulis, and W. Durante YC-1 Stimulates the Expression of Gaseous Monoxide-Generating Enzymes in Vascular Smooth Muscle Cells Mol. Pharmacol., January 1, 2009; 75(1): 208 - 217. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. Russo, P. Del Mese, G. Doronzo, L. Mattiello, M. Viretto, A. Bosia, G. Anfossi, and M. Trovati Resistance to the Nitric Oxide/Cyclic Guanosine 5'-Monophosphate/Protein Kinase G Pathway in Vascular Smooth Muscle Cells from the Obese Zucker Rat, a Classical Animal Model of Insulin Resistance: Role of Oxidative Stress Endocrinology, April 1, 2008; 149(4): 1480 - 1489. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X. Wang, J. L. Pluznick, D. C. Settles, and S. C. Sansom Association of VASP with TRPC4 in PKG-mediated inhibition of the store-operated calcium response in mesangial cells Am J Physiol Renal Physiol, December 1, 2007; 293(6): F1768 - F1776. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Tamada, M. Nara, H. Kanatsuka, M. Nagaoka, R. Koshida, G. Tamura, and T. Hattori A Potentiating Effect of Endogenous NO in the Physiologic Secretion from Airway Submucosal Glands Am. J. Respir. Cell Mol. Biol., September 1, 2007; 37(3): 357 - 365. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. E. Teixeira, F. B. M. Priviero, and R. C. Webb Molecular Mechanisms Underlying Rat Mesenteric Artery Vasorelaxation Induced by the Nitric Oxide-Independent Soluble Guanylyl Cyclase Stimulators BAY 41-2272 [5-Cyclopropyl-2-[1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]pyrimidin-4-ylamine] and YC-1 [3-(5'-Hydroxymethyl-2'-furyl)-1-benzyl Indazole] J. Pharmacol. Exp. Ther., April 1, 2006; 317(1): 258 - 266. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. J. Busch, H. Liu, A. R. Graveline, and K. D. Bloch Nitric oxide induces phosphodiesterase 4B expression in rat pulmonary artery smooth muscle cells Am J Physiol Lung Cell Mol Physiol, April 1, 2006; 290(4): L747 - L753. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Hofmann, R. Feil, T. Kleppisch, and J. Schlossmann Function of cGMP-Dependent Protein Kinases as Revealed by Gene Deletion Physiol Rev, January 1, 2006; 86(1): 1 - 23. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. P.Y. Lee, D. D. Mruk, C.-h. Wong, and C. Y. Cheng Regulation of Sertoli-Germ Cell Adherens Junction Dynamics in the Testis Via the Nitric Oxide Synthase (NOS)/cGMP/Protein Kinase G (PRKG)/{beta}-Catenin (CATNB) Signaling Pathway: An In Vitro and In Vivo Study Biol Reprod, September 1, 2005; 73(3): 458 - 471. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. G. Werner, V. Godfrey, R. R. Arnold, G. L. Featherstone, D. Bender, J. Schlossmann, M. Schiemann, F. Hofmann, and K. B. Pryzwansky Neutrophil Dysfunction in Guanosine 3',5'-Cyclic Monophosphate-Dependent Protein Kinase I-Deficient Mice J. Immunol., August 1, 2005; 175(3): 1919 - 1929. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 U. Walter and S. Gambaryan Roles of cGMP/cGMP-dependent protein kinase in platelet activation Blood, October 15, 2004; 104(8): 2609 - 2609. [Full Text] [PDF]
|
|
|
|
 |
|
 L. Chen, G. Daum, K. Chitaley, S. A. Coats, D. F. Bowen-Pope, M. Eigenthaler, N. R. Thumati, U. Walter, and A. W. Clowes Vasodilator-Stimulated Phosphoprotein Regulates Proliferation and Growth Inhibition by Nitric Oxide in Vascular Smooth Muscle Cells Arterioscler. Thromb. Vasc. Biol., August 1, 2004; 24(8): 1403 - 1408. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. S. Taylor, C. Okwuchukwuasanya, C. K. Nickl, W. Tegge, J. E. Brayden, and W. R. G. Dostmann Inhibition of cGMP-Dependent Protein Kinase by the Cell-Permeable Peptide DT-2 Reveals a Novel Mechanism of Vasoregulation Mol. Pharmacol., May 1, 2004; 65(5): 1111 - 1119. [Abstract] [Full Text]
|
|
|
|
 |
|
 K. Kohler, D. Louvard, and A. Zahraoui Rab13 regulates PKA signaling during tight junction assembly J. Cell Biol., April 26, 2004; 165(2): 175 - 180. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Gambaryan, J. Geiger, U. R. Schwarz, E. Butt, A. Begonja, A. Obergfell, and U. Walter Potent inhibition of human platelets by cGMP analogs independent of cGMP-dependent protein kinase Blood, April 1, 2004; 103(7): 2593 - 2600. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H.-Y. Kwan, Y. Huang, and X. Yao Regulation of canonical transient receptor potential isoform 3 (TRPC3) channel by protein kinase G PNAS, February 24, 2004; 101(8): 2625 - 2630. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Randriamboavonjy, J. Schrader, R. Busse, and I. Fleming Insulin Induces the Release of Vasodilator Compounds From Platelets by a Nitric Oxide-G Kinase-VAMP-3-dependent Pathway J. Exp. Med., February 2, 2004; 199(3): 347 - 356. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. H. H. Chan, K.-F. Chang, C.-C. Ou, and J. Y. H. Chan Nitric Oxide Regulates c-fos Expression in Nucleus Tractus Solitarii Induced by Baroreceptor Activation via cGMP-Dependent Protein Kinase and cAMP Response Element-Binding Protein Phosphorylation Mol. Pharmacol., February 1, 2004; 65(2): 319 - 325. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Sengenes, A. Bouloumie, H. Hauner, M. Berlan, R. Busse, M. Lafontan, and J. Galitzky Involvement of a cGMP-dependent Pathway in the Natriuretic Peptide-mediated Hormone-sensitive Lipase Phosphorylation in Human Adipocytes J. Biol. Chem., December 5, 2003; 278(49): 48617 - 48626. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. B. Pilz and D. E. Casteel Regulation of Gene Expression by Cyclic GMP Circ. Res., November 28, 2003; 93(11): 1034 - 1046. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Feil, S. M. Lohmann, H. de Jonge, U. Walter, and F. Hofmann Cyclic GMP-Dependent Protein Kinases and the Cardiovascular System: Insights From Genetically Modified Mice Circ. Res., November 14, 2003; 93(10): 907 - 916. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. Wolfsgruber, S. Feil, S. Brummer, O. Kuppinger, F. Hofmann, and R. Feil A proatherogenic role for cGMP-dependent protein kinase in vascular smooth muscle cells PNAS, November 11, 2003; 100(23): 13519 - 13524. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Munzel, R. Feil, A. Mulsch, S. M. Lohmann, F. Hofmann, and U. Walter Physiology and Pathophysiology of Vascular Signaling Controlled by Cyclic Guanosine 3',5'-Cyclic Monophosphate-Dependent Protein Kinase Circulation, November 4, 2003; 108(18): 2172 - 2183. [Full Text] [PDF]
|
|
|
|
|
|
R. Feil, J. Hartmann, C. Luo, W. Wolfsgruber, K. Schilling, S. Feil, J. J. Barski, M. Meyer, A. Konnerth, C. I. De Zeeuw, et al. Impairment of LTD and cerebellar learning by Purkinje cell-specific ablation of cGMP-dependent protein kinase I J. Cell Biol., October 27, 2003; 163(2): 295 - 302. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Gambaryan, E. Butt, K. Marcus, M. Glazova, A. Palmetshofer, G. Guillon, and A. Smolenski cGMP-dependent Protein Kinase Type II Regulates Basal Level of Aldosterone Production by Zona Glomerulosa Cells without Increasing Expression of the Steroidogenic Acute Regulatory Protein Gene J. Biol. Chem., August 8, 2003; 278(32): 29640 - 29648. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. P. Y. Lee and C. Yan Cheng Regulation of Sertoli Cell Tight Junction Dynamics in the Rat Testis via the Nitric Oxide Synthase/Soluble Guanylate Cyclase/3',5'-Cyclic Guanosine Monophosphate/Protein Kinase G Signaling Pathway: an in Vitro Study Endocrinology, July 1, 2003; 144(7): 3114 - 3129. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Li, J. Ajdic, M. Eigenthaler, and X. Du A predominant role for cAMP-dependent protein kinase in the cGMP-induced phosphorylation of vasodilator-stimulated phosphoprotein and platelet inhibition in humans Blood, June 1, 2003; 101(11): 4423 - 4429. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Schmidt, M. Werner, P. A. Heppenstall, M. Henning, M. I. More, S. Kuhbandner, G. R. Lewin, F. Hofmann, R. Feil, and F. G. Rathjen cGMP-mediated signaling via cGKI{alpha} is required for the guidance and connectivity of sensory axons J. Cell Biol., November 7, 2002; 159(3): 489 - 498. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Zeng, Y. Liu, and D. M. Templeton Ca2+/calmodulin-dependent and cAMP-dependent kinases in induction of c-fos in human mesangial cells Am J Physiol Renal Physiol, November 1, 2002; 283(5): F888 - F894. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. K. Taylor, R. Ahmed, M. Begley, and M. D. Uhler Autoinhibition and Isoform-specific Dominant Negative Inhibition of the Type II cGMP-dependent Protein Kinase J. Biol. Chem., September 27, 2002; 277(40): 37242 - 37253. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. R. Hassock, M. X. Zhu, C. Trost, V. Flockerzi, and K. S. Authi Expression and role of TRPC proteins in human platelets: evidence that TRPC6 forms the store-independent calcium entry channel Blood, September 26, 2002; 100(8): 2801 - 2811. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. den Dekker, G. Gorter, J. W. M. Heemskerk, and J.-W. N. Akkerman Development of Platelet Inhibition by cAMP during Megakaryocytopoiesis J. Biol. Chem., August 2, 2002; 277(32): 29321 - 29329. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Carabelli, M. D'Ascenzo, E. Carbone, and C. Grassi Nitric oxide inhibits neuroendocrine CaV1 L-channel gating via cGMP-dependent protein kinase in cell-attached patches of bovine chromaffin cells J. Physiol., June 1, 2002; 541(2): 351 - 366. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Feil, N. Gappa, M. Rutz, J. Schlossmann, C. R. Rose, A. Konnerth, S. Brummer, S. Kuhbandner, and F. Hofmann Functional Reconstitution of Vascular Smooth Muscle Cells With cGMP-Dependent Protein Kinase I Isoforms Circ. Res., May 31, 2002; 90(10): 1080 - 1086. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Bader, E. Butt, A. Palmetshofer, U. Walter, T. Jarchau, and P. Drueckesl A cGMP-Dependent Protein Kinase Assay for High Throughput Screening Based on Time-Resolved Fluorescence Resonance Energy Transfer J Biomol Screen, August 1, 2001; 6(4): 255 - 264. [Abstract] [PDF]
|
|
|
|
|
|
T. A. Fischer, A. Palmetshofer, S. Gambaryan, E. Butt, C. Jassoy, U. Walter, S. Sopper, and S. M. Lohmann Activation of cGMP-dependent Protein Kinase Ibeta Inhibits Interleukin 2 Release and Proliferation of T Cell Receptor-stimulated Human Peripheral T Cells J. Biol. Chem., February 16, 2001; 276(8): 5967 - 5974. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |