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Related Experiment Videos

Intracellular acidosis activates c-Src

Y Yamaji1, H Tsuganezawa, O W Moe

  • 1Department of Internal Medicine, The University of Texas Southwestern Medical Center, Dallas 75235, USA.

The American Journal of Physiology
|March 1, 1997
PubMed
Summary

Acidosis activates the c-Src protein tyrosine kinase pathway. This study shows that decreased intracellular pH rapidly activates c-Src, a novel finding with implications for cellular responses to acidosis.

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Renal Physiology

Background:

  • Acidosis, a condition of excess acid in the body, can impact cellular function.
  • Protein tyrosine kinases are crucial signaling enzymes involved in various cellular processes.
  • The specific role of acidosis in activating protein tyrosine kinase pathways remains largely unexplored.

Purpose of the Study:

  • To investigate whether acidosis triggers the activation of protein tyrosine kinase pathways.
  • To determine the effect of altered intracellular pH on c-Src kinase activity.

Main Methods:

  • Incubation of MCT cells (a renal proximal tubule cell line) in acidic media.
  • Measurement of phosphotyrosine content in cytosolic proteins.
  • Assay of c-Src kinase activity.

Related Experiment Videos

  • Manipulation of intracellular pH using sodium propionate, NH4Cl prepulse, and nigericin.
  • Use of sodium orthovanadate, a protein tyrosine phosphatase inhibitor.

    • Main Results:

    • Acidic media incubation increased phosphotyrosine content in 60-70 kDa and 120 kDa cytosolic proteins.
    • Media acidification led to a rapid, twofold increase in c-Src activity.
    • Significant c-Src activation was observed with minor changes in extracellular and intracellular pH.
    • Maneuvers decreasing intracellular pH activated c-Src, independent of extracellular pH changes.
    • Sodium orthovanadate inhibited c-Src activation by acidification but not protein tyrosine phosphorylation.

    Conclusions:

    • Decreased intracellular pH is a direct activator of c-Src kinase.
    • Acid-induced activation of c-Src represents a novel signaling mechanism.
    • This finding may be relevant to understanding cellular responses to acidosis in various physiological contexts.