In vitro receptor binding properties of a "painless" NGF mutein, linked to hereditary sensory autonomic neuropathy type V.

Abstract

Nerve Growth Factor (NGF) signalling is mediated by the TrkA and p75NTR receptors. Besides its neurotrophic and survival activities, NGF displays a potent pro-nociceptive activity. Recently, a missense point mutation was found in the NGFB gene (C661T, leading to the aminoacid substitution R100W) of individuals affected by a form of hereditary loss of pain perception (hereditary sensory and autonomic neuropathy type V, HSAN V). In order to gain insights into the functional consequences of the HSAN V NGF mutation, two sets of hNGFR100 mutants were expressed in Escherichia coli and purified, as mature NGF or proNGF, for in vitro receptor binding studies. Here, we show by Surface Plasmon Resonance analysis that the R100 mutation selectively disrupts binding of hNGF to p75NTR receptor, to an extent which depends on the substituting residue at position 100, while the affinity of hNGFR100 mutants for TrkA receptor is not affected. As for unprocessed hproNGF, the binding of the R100 variants to p75NTR receptor shows only a limited impairment, showing that the impact of the R100 mutation on p75NTR receptor binding is greater in the context of mature, processed hNGF. These results provide a basis for elucidating the mechanisms underlying the clinical manifestations of HSAN V patients, and provide a basis for the development of "painless" hNGF molecules with therapeutic potential.

• Authors: Sonia Covaceuszach, Simona Capsoni, Sara Marinelli, Flaminia Pavone, Marcello Ceci, Gabriele Ugolini, Domenico Vignone, Gianluca Amato, Francesca Paoletti, Doriano Lamba, Antonino Cattaneo
• Journal: Biochemical and biophysical research communications (Biochem Biophys Res Commun) Vol. 391 Issue 1 Pg. 824-9 (Jan 01 2010) ISSN: 1090-2104 [Electronic] United States
• PMID: 19945432 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright 2009 Elsevier Inc. All rights reserved.

Chemical References

• NGF protein, human
• NGFR protein, human
• Nerve Tissue Proteins
• Receptors, Nerve Growth Factor
• Nerve Growth Factor
• Receptor, trkA

Topics

• Escherichia coli (genetics, metabolism)
• Hereditary Sensory and Autonomic Neuropathies (genetics, metabolism)
• Humans
• Mutation
• Nerve Growth Factor (chemistry, genetics, metabolism)
• Nerve Tissue Proteins (metabolism)
• Pain Insensitivity, Congenital (genetics, metabolism)
• Protein Conformation
• Receptor, trkA (metabolism)
• Receptors, Nerve Growth Factor (metabolism)
• Surface Plasmon Resonance