Human ProNGF: biological effects and binding profiles at TrkA, P75NTR and sortilin

J Neurochem. 2008 Nov;107(4):1124-35. doi: 10.1111/j.1471-4159.2008.05698.x. Epub 2008 Sep 21.

Abstract

Nerve growth factor (NGF) promotes cell survival via binding to the tyrosine kinase receptor A (TrkA). Its precursor, proNGF, binds to p75(NTR) and sortilin receptors to initiate apoptosis. Current disagreement exists over whether proNGF acts neurotrophically following binding to TrkA. As in Alzheimer's disease the levels of proNGF increase and TrkA decrease, it is important to clarify the properties of proNGF. Here, wild-type and cleavage-resistant mutated forms (M) of proNGF were engineered and their binding characteristics determined. M-proNGF and NGF bound to p75(NTR) with similar affinities, whilst M-proNGF had a lower affinity than NGF for TrkA. M-proNGF behaved neurotrophically, albeit less effectively than NGF. M-proNGF addition resulted in phosphorylation of TrkA and ERK1/2, and in PC12 cells elicited neurite outgrowth and supported cell survival. Conversely, M-proNGF addition to cultured cortical neurons initiated caspase 3 cleavage. Importantly, these biological effects were shown to be mediated by unprocessed M-proNGF. Surprisingly, binding of the pro region alone to TrkA, at a site other than that of NGF, caused TrkA and ERK1/2 phosphorylation. Our data show that M-proNGF stimulates TrkA to a lesser degree than NGF, suggesting that in Alzheimer brain the increased proNGF : NGF and p75(NTR) : TrkA ratios may permit apoptotic effects to predominate over neurotrophic effects.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Vesicular Transport / metabolism*
  • Animals
  • Animals, Newborn
  • Caspase 3 / metabolism
  • Cell Survival
  • Cells, Cultured
  • Cerebral Cortex / cytology
  • Humans
  • Mitogen-Activated Protein Kinase 3 / metabolism
  • Mutation / physiology
  • Nerve Growth Factor / genetics
  • Nerve Growth Factor / metabolism*
  • Nerve Tissue Proteins
  • Neurites / metabolism
  • Neurons / drug effects
  • Neurons / metabolism*
  • Phosphorylation
  • Protein Binding
  • Protein Precursors / genetics
  • Protein Precursors / metabolism*
  • Radioligand Assay / methods
  • Rats
  • Receptor, trkA / metabolism*
  • Receptors, Growth Factor
  • Receptors, Nerve Growth Factor / metabolism*
  • Recombinant Proteins
  • Signal Transduction / physiology
  • Time Factors

Substances

  • Adaptor Proteins, Vesicular Transport
  • Nerve Tissue Proteins
  • Protein Precursors
  • Receptors, Growth Factor
  • Receptors, Nerve Growth Factor
  • Recombinant Proteins
  • pro-nerve growth factor, human
  • Ngfr protein, rat
  • Nerve Growth Factor
  • Receptor, trkA
  • Mitogen-Activated Protein Kinase 3
  • Caspase 3
  • sortilin