![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
![[Contents]](/icons/banner/tocACT.gif){kind=icon}
|
|
|
|
|
||
Vol. 17, Issue 1, 317-326, January 2006
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
* Departments of Pharmacology and Medicine, Medical University of Ohio, Toledo, OH 43614;
Department of Physiology, Cornell University, Weill Medical College, New York, NY 10021
Submitted August 9, 2005;
Revised September 27, 2005;
Accepted October 21, 2005
Monitoring Editor: Guido Guidotti
We have shown that ouabain activates Src, resulting in subsequent tyrosine phosphorylation of multiple effectors. Here, we tested if the Na+/K+-ATPase and Src can form a functional signaling complex. In LLC-PK1 cells the Na+/K+-ATPase and Src colocalized in the plasma membrane. Fluorescence resonance energy transfer analysis indicated that both proteins were in close proximity, suggesting a direct interaction. GST pulldown assay showed a direct, ouabain-regulated, and multifocal interaction between the 
1 subunit of Na+/K+-ATPase and Src. Although the interaction between the Src kinase domain and the third cytosolic domain (CD3) of 1 is regulated by ouabain, the Src SH3SH2 domain binds to the second cytosolic domain constitutively. Functionally, binding of Src to either the Na+/K+-ATPase or GST-CD3 inhibited Src activity. Addition of ouabain, but not vanadate, to the purified Na+/K+-ATPase/Src complex freed the kinase domain and restored the Src activity. Consistently, exposure of intact cells to ouabain apparently increased the distance between the Na+/K+-ATPase and Src. Concomitantly, it also stimulated tyrosine phosphorylation of the proteins that are associated with the Na+/K+-ATPase. These new findings illustrate a novel molecular mechanism of signal transduction involving the interaction of a P-type ATPase and a nonreceptor tyrosine kinase.
Abbreviations used: BRET, bioluminescence resonance energy transfer; ECFP, enhanced cyan fluorescent protein; EYFP, enhanced yellow fluorescent protein; FRET, fluorescence resonance energy transfer; GST, glutathione-S-transferase; IB, immunoblotting; IP, immunoprecipitation; PKE, purified pig kidney enzyme (Na+/K+-ATPase); PP2, 4-amino-5-[4-chlorophenyl]-7-[t-butyl]pyrazolo[3,4-d]pyrimidine; PP3, 4-amino-7-phenylpyrazol[3,4]pyromidine; RIPA, radioimmunoprecipitation.
These authors contributed equally to this work.
Address correspondence to: Zi-Jian Xie (zxie{at}meduohio.edu).
This article has been cited by other articles:
|
|
 |
|
  J. Tian and Z.-j. Xie The Na-K-ATPase and Calcium-Signaling Microdomains Physiology, August 1, 2008; 23(4): 205 - 211. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X. Liu, Z. Spicarova, S. Rydholm, J. Li, H. Brismar, and A. Aperia Ankyrin B Modulates the Function of Na,K-ATPase/Inositol 1,4,5-Trisphosphate Receptor Signaling Microdomain J. Biol. Chem., April 25, 2008; 283(17): 11461 - 11468. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. R. Yingst, T. M. Doci, K. J. Massey, N. F. Rossi, E. Rucker, and R. R. Mattingly Angiotensin II stimulates elution of Na-K-ATPase from a digoxin-affinity column by increasing the kinetic response to ligands that trigger the decay of E2-P Am J Physiol Renal Physiol, April 1, 2008; 294(4): F990 - F1000. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Fujii, Y. Takahashi, Y. Itomi, K. Fujita, M. Morii, Y. Tabuchi, S. Asano, K. Tsukada, N. Takeguchi, and H. Sakai K+-Cl- Cotransporter-3a Up-regulates Na+,K+-ATPase in Lipid Rafts of Gastric Luminal Parietal Cells J. Biol. Chem., March 14, 2008; 283(11): 6869 - 6877. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. M. Lynch, C. S. Weber, K. D. Nullmeyer, E. D. W. Moore, and R. J. Paul Clearance of store-released Ca2+ by the Na+-Ca2+ exchanger is diminished in aortic smooth muscle from Na+-K+-ATPase {alpha}2-isoform gene-ablated mice Am J Physiol Heart Circ Physiol, March 1, 2008; 294(3): H1407 - H1416. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Cai, L. Wu, W. Qu, D. Malhotra, Z. Xie, J. I. Shapiro, and J. Liu Regulation of apical NHE3 trafficking by ouabain-induced activation of the basolateral Na+-K+-ATPase receptor complex Am J Physiol Cell Physiol, February 1, 2008; 294(2): C555 - C563. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Chen, T. Cai, C. Yang, D. A. Turner, D. R. Giovannucci, and Z. Xie Regulation of Inositol 1,4,5-Trisphosphate Receptor-mediated Calcium Release by the Na/K-ATPase in Cultured Renal Epithelial Cells J. Biol. Chem., January 11, 2008; 283(2): 1128 - 1136. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Liu, X. Zhao, S. V. Pierre, and A. Askari Association of PI3K-Akt signaling pathway with digitalis-induced hypertrophy of cardiac myocytes Am J Physiol Cell Physiol, November 1, 2007; 293(5): C1489 - C1497. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Edwards and T. L. Pallone Ouabain modulation of cellular calcium stores and signaling Am J Physiol Renal Physiol, November 1, 2007; 293(5): F1518 - F1532. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. J. Khundmiri, V. Amin, J. Henson, J. Lewis, M. Ameen, M. J. Rane, and N. A. Delamere Ouabain stimulates protein kinase B (Akt) phosphorylation in opossum kidney proximal tubule cells through an ERK-dependent pathway Am J Physiol Cell Physiol, September 1, 2007; 293(3): C1171 - C1180. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. Schoner and G. Scheiner-Bobis Endogenous and exogenous cardiac glycosides: their roles in hypertension, salt metabolism, and cell growth Am J Physiol Cell Physiol, August 1, 2007; 293(2): C509 - C536. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Edwards and T. L. Pallone Modification of cytosolic calcium signaling by subplasmalemmal microdomains Am J Physiol Renal Physiol, June 1, 2007; 292(6): F1827 - F1845. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Liang, J. Tian, L. Liu, S. Pierre, J. Liu, J. Shapiro, and Z.-J. Xie Identification of a Pool of Non-pumping Na/K-ATPase J. Biol. Chem., April 6, 2007; 282(14): 10585 - 10593. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A.-F. Ruaud and J.-L. Bessereau The P-type ATPase CATP-1 is a novel regulator of C. elegans developmental timing that acts independently of its predicted pump function Development, March 1, 2007; 134(5): 867 - 879. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Inserte Triggering of cardiac preconditioning through Na+/K+-ATPase Cardiovasc Res, February 1, 2007; 73(3): 446 - 447. [Full Text] [PDF]
|
|
|
|
|
|
S. V. Pierre, C. Yang, Z. Yuan, J. Seminerio, C. Mouas, K. D. Garlid, P. Dos-Santos, and Z. Xie Ouabain triggers preconditioning through activation of the Na+,K+-ATPase signaling cascade in rat hearts Cardiovasc Res, February 1, 2007; 73(3): 488 - 496. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. A. Rajasekaran, S. P. Barwe, J. Gopal, S. Ryazantsev, E. E. Schneeberger, and A. K. Rajasekaran Na-K-ATPase regulates tight junction permeability through occludin phosphorylation in pancreatic epithelial cells Am J Physiol Gastrointest Liver Physiol, January 1, 2007; 292(1): G124 - G133. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
O. Kotova, L. Al-Khalili, S. Talia, C. Hooke, O. V. Fedorova, A. Y. Bagrov, and A. V. Chibalin Cardiotonic Steroids Stimulate Glycogen Synthesis in Human Skeletal Muscle Cells via a Src- and ERK1/2-dependent Mechanism J. Biol. Chem., July 21, 2006; 281(29): 20085 - 20094. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. Larre, A. Ponce, R. Fiorentino, L. Shoshani, R. G. Contreras, and M. Cereijido Contacts and cooperation between cells depend on the hormone ouabain PNAS, July 18, 2006; 103(29): 10911 - 10916. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Liang, T. Cai, J. Tian, W. Qu, and Z.-J. Xie Functional Characterization of Src-interacting Na/K-ATPase Using RNA Interference Assay J. Biol. Chem., July 14, 2006; 281(28): 19709 - 19719. [Abstract] [Full Text] [PDF]
|
|
