SMPL synaptic membranes

Nanodisc-mediated synaptic membrane mimetics expand the toolkit for drug discovery and the molecular cell biology of synapses

Riana Lo Bu, Jeffrey Clark, Thomas DiChiara, Stephen G. Sligar, William L Klein*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

We describe in detail how proteins of synaptic membranes can be extracted and reconstituted within “Nanodiscs” which renders them soluble in aqueous solutions. Each Nanodisc is a nanometer-scaled lipid bilayer containing an individual membrane protein. The method can provide a soluble membrane protein library (SMPL) from any tissue fraction, notably from the standpoint of this treatise, from synaptosome plasma membranes (SMPL synaptic membranes). Nanodiscs facilitate investigations of membrane proteins that are difficult to tease apart from protein-protein complexes while at the same time maintaining conformational integrity. The procedure captures virtually all membrane proteins in a manner that preserves their activities in soluble form. The usefulness of SMPL synaptic membranes is exemplified by experiments that concern the specific binding of Alzheimer’s-related amyloid β oligomers (AβOs). AβO binding to SMPL synaptic membranes is saturable, has the same high affinity found in cell-based assays, and is mediated adventitiously by particular proteins. Immunoaffinity isolation experiments have identified one AβO toxin “receptor” as the NaK ATPase α3 (NKAα3). For drug discovery, recombinant membrane proteins in Nanodiscs can be readily coupled to high-throughput screening platforms; SMPL synaptic membranes can also be readily coupled and used for unbiased high-throughput screening of unknown targets. SMPL synaptic membranes, and analogous organelle libraries, offer new resources for the CNS membrane protein toolkit with a wide array of applications to synapse molecular cell biology and drug discovery.

Original languageEnglish (US)
Title of host publicationNeuromethods
PublisherHumana Press Inc
Pages227-250
Number of pages24
DOIs
StatePublished - Jan 1 2018

Publication series

NameNeuromethods
Volume141
ISSN (Print)0893-2336
ISSN (Electronic)1940-6045

Fingerprint

Cytology
Synaptic Membranes
Molecular biology
Drug Discovery
Synapses
Cell Biology
Molecular Biology
Membrane Proteins
Membranes
Oligomers
Amyloid
Screening
Proteins
Throughput
Lipid bilayers
Synaptosomes
Lipid Bilayers
Cell membranes
Recombinant Proteins
Organelles

Keywords

  • Abeta oligomers
  • Alzheimer’s
  • Nanodisc
  • Receptor
  • Synapse
  • Synaptic membrane

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)
  • Psychiatry and Mental health

Cite this

Lo Bu, Riana ; Clark, Jeffrey ; DiChiara, Thomas ; Sligar, Stephen G. ; Klein, William L. / SMPL synaptic membranes : Nanodisc-mediated synaptic membrane mimetics expand the toolkit for drug discovery and the molecular cell biology of synapses. Neuromethods. Humana Press Inc, 2018. pp. 227-250 (Neuromethods).
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SMPL synaptic membranes : Nanodisc-mediated synaptic membrane mimetics expand the toolkit for drug discovery and the molecular cell biology of synapses. / Lo Bu, Riana; Clark, Jeffrey; DiChiara, Thomas; Sligar, Stephen G.; Klein, William L.

Neuromethods. Humana Press Inc, 2018. p. 227-250 (Neuromethods; Vol. 141).

Research output: Chapter in Book/Report/Conference proceedingChapter

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