Characterizing nerve growth factor-p75(NTR) interactions and small molecule inhibition using surface plasmon resonance spectroscopy

Kristen S A Sheffield; Allison E Kennedy; John A Scott; Gregory M Ross

doi:10.1016/j.ab.2015.09.019

Characterizing nerve growth factor-p75(NTR) interactions and small molecule inhibition using surface plasmon resonance spectroscopy

Anal Biochem. 2016 Jan 15:493:21-6. doi: 10.1016/j.ab.2015.09.019. Epub 2015 Oct 3.

Authors

Kristen S A Sheffield¹, Allison E Kennedy², John A Scott³, Gregory M Ross⁴

Affiliations

¹ Department of Biology, Laurentian University, Sudbury, Ontario P3E 2C6, Canada.
² Department of Biomolecular Sciences, Laurentian University, Sudbury, Ontario P3E 2C6, Canada.
³ Department of Biomolecular Sciences, Laurentian University, Sudbury, Ontario P3E 2C6, Canada; Bharti School of Engineering, Laurentian University, Sudbury, Ontario P3E 2C6, Canada; Northern Ontario School of Medicine, Sudbury, Ontario P3E 2C6, Canada.
⁴ Department of Biology, Laurentian University, Sudbury, Ontario P3E 2C6, Canada; Department of Biomolecular Sciences, Laurentian University, Sudbury, Ontario P3E 2C6, Canada; Northern Ontario School of Medicine, Sudbury, Ontario P3E 2C6, Canada. Electronic address: gross@nosm.ca.

PMID: 26435172
DOI: 10.1016/j.ab.2015.09.019

Abstract

Nerve growth factor (NGF) is critical for the proliferation, differentiation, and survival of neurons through its binding to the p75(NTR) and TrkA receptors. Dysregulation of NGF has been implicated in several pathologies, including neurodegeneration (i.e., Parkinson's and Alzheimer's diseases) and both inflammatory and neuropathic pain states. Therefore, small molecule inhibitors that block NGF-receptor interactions have significant therapeutic potential. Small molecule antagonists ALE-0540, PD90780, Ro 08-2750, and PQC 083 have all been reported to inhibit NGF from binding the TrkA receptor. Interestingly, the characterization of the ability of these molecules to block NGF-p75(NTR) interactions has not been performed. In addition, the inhibitory action of these molecules has never been evaluated using surface plasmon resonance (SPR) spectroscopy, which has been proven to be highly useful in drug discovery applications. In the current study, we used SPR biosensors to characterize the binding of NGF to the p75(NTR) receptor in addition to characterizing the inhibitory potential of the known NGF antagonists. The results of this study provide the first evaluation of the ability of these compounds to block NGF binding to p75(NTR) receptor. In addition, only PD90780 was effective at inhibiting the interaction of NGF with p75(NTR), suggesting receptor selectivity between known NGF inhibitors.

Keywords: Biosensor; NGF inhibition; Nerve growth factor; Surface plasmon resonance; TrkA; p75(NTR).

MeSH terms

Flavins
Heterocyclic Compounds, 3-Ring / chemistry
Heterocyclic Compounds, 3-Ring / pharmacology
Humans
Nerve Growth Factor / antagonists & inhibitors
Nerve Growth Factor / metabolism*
Protein Binding / drug effects
Protein Interaction Maps / drug effects*
Pteridines / chemistry
Pteridines / pharmacology
Receptor, Nerve Growth Factor / antagonists & inhibitors
Receptor, Nerve Growth Factor / metabolism*
Receptor, trkA / antagonists & inhibitors
Receptor, trkA / metabolism
Small Molecule Libraries / chemistry*
Small Molecule Libraries / pharmacology*
Surface Plasmon Resonance / methods*

Substances

2,3,4,10-tetrahydro-7,10-dimethyl-2,4-dioxobenzo(g)pteridine
Flavins
Heterocyclic Compounds, 3-Ring
Pteridines
Receptor, Nerve Growth Factor
Small Molecule Libraries
ALE 0540
Nerve Growth Factor
Receptor, trkA