Binding of Src to Na+/K+-ATPase forms a functional signaling complex. Academic Article uri icon

Overview

abstract

  • 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.

publication date

  • November 2, 2005

Research

keywords

  • Signal Transduction
  • Sodium-Potassium-Exchanging ATPase
  • src-Family Kinases

Identity

PubMed Central ID

  • PMC1345669

Scopus Document Identifier

  • 30044451651

Digital Object Identifier (DOI)

  • 10.1091/mbc.e05-08-0735

PubMed ID

  • 16267270

Additional Document Info

volume

  • 17

issue

  • 1