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Evidence for communication between nerve growth factor and protein tyrosine phosphorylation

N Gómez1, N K Tonks, C Morrison

  • 1Department of Biochemistry, University of Dundee, UK.

FEBS Letters
|October 1, 1990
PubMed

Insights

Nerve growth factor (NGF) activates myelin basic protein (MBP) kinases in PC12 cells. These kinases, regulated by protein tyrosine and serine/threonine phosphatases, link NGF signaling to cellular phosphorylation.

Area of Science:

  • Neuroscience
  • Cell Biology
  • Biochemistry

Background:

  • Nerve growth factor (NGF) is crucial for neuronal development and survival.
  • PC12 cells are a widely used model system for studying NGF signaling pathways.
  • Understanding the early molecular events downstream of NGF is key to deciphering neuronal communication.

Purpose of the Study:

  • To identify and characterize the early kinase activities activated by NGF in PC12 cells.
  • To investigate the role of protein phosphatases in regulating these NGF-induced kinase activities.
  • To elucidate the mechanism by which NGF signaling influences intracellular protein phosphorylation.

Main Methods:

  • PC12 cells were stimulated with NGF.
  • Myelin basic protein (MBP) kinase activities were measured.
  • Enzymes were resolved using Mono Q chromatography.
  • Phosphorylation sites were analyzed.
  • Enzyme inactivation assays were performed using specific phosphatases (CD45, PP2A) and inhibitors (vanadate, okadaic acid).

Main Results:

  • NGF stimulation rapidly activated two distinct MBP kinase activities (>10-fold within 5 min).
  • Both kinases phosphorylated MBP on threonine residues.
  • The activated kinases were inactivated by both a protein tyrosine phosphatase (CD45) and a serine/threonine phosphatase (PP2A).
  • Vanadate blocked CD45 inactivation, and okadaic acid blocked PP2A inactivation, confirming phosphatase activity.

Conclusions:

  • NGF rapidly activates specific MBP kinase activities in PC12 cells.
  • These kinases are subject to regulation by both tyrosine and serine/threonine phosphatases.
  • The activation of MBP-kinases represents a critical signaling step connecting NGF stimulation to intracellular protein tyrosine phosphorylation.

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