Nanoscale Synaptic Membrane Mimetic Allows Unbiased High Throughput Screen That Targets Binding Sites for Alzheimer’s-Associated Aβ Oligomers

Wilcox, Kyle C.; Marunde, Matthew R.; Das, Aditi; Velasco, Pauline T.; Kuhns, Benjamin D.; Marty, Michael T.; Haoming, Jiang; Luan, Chi Hao; Sligar, Stephen G.; Klein, William L.

doi:10.1371/journal.pone.0125263

Cited by 30 publications

(36 citation statements)

References 68 publications

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“…111 Synaptic plasma membranes from rat cerebral cortex were used to prepare synaptosomes, which were solubilized in the detergent DDM and used for Nanodisc assembly with MSP1E3D1 and POPC. After removal of detergent on hydrophobic beads, Nanodiscs were purified by Ni-NTA affinity chromatography using the His-tag on MSP.…”

Section: Nanodisc Applications In Biotechnology and Medicinementioning

confidence: 99%

Nanodiscs in Membrane Biochemistry and Biophysics

2017

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Membrane proteins play a most important part in metabolism, signaling, cell motility, transport, development, and many other biochemical and biophysical processes which constitute fundamentals of life on molecular level. Detailed understanding of these processes is necessary for the progress of life sciences and biomedical applications. Nanodiscs provide a new and powerful tool for a broad spectrum of biochemical and biophysical studies of membrane proteins and are commonly acknowledged as an optimal membrane mimetic system which provides control over size, composition and specific functional modifications on nanometer scale. In this review we attempted to combine a comprehensive list of various applications of Nanodisc technology with systematic analysis of the most attractive features of this system and advantages provided by Nanodiscs for structural and mechanistic studies of membrane proteins.

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Section: Nanodisc Applications In Biotechnology and Medicinementioning

confidence: 99%

Nanodiscs in Membrane Biochemistry and Biophysics

2017

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“…Not only Ca 2+ but cAMP levels [24, 156] could be disrupted. It is intriguing that once neurons have reached a fully differentiated state, PrP can enzymatically be removed with little or no impact on AβO accumulation in hot spots and synapses [180]. …”

Section: Transduction Mechanismmentioning

confidence: 99%

Amyloid β oligomers in Alzheimer’s disease pathogenesis, treatment, and diagnosis

2015

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Protein aggregation is common to dozens of diseases including prionoses, diabetes, Parkinson’s and Alzheimer’s. Over the past 15 years, there has been a paradigm shift in understanding the structural basis for these proteinopathies. Precedent for this shift has come from investigation of soluble Aβ oligomers (AβOs), toxins now widely regarded as instigating neuron damage leading to Alzheimer’s dementia. Toxic AβOs accumulate in AD brain and constitute long-lived alternatives to the disease-defining Aβ fibrils deposited in amyloid plaques. Key experiments using fibril-free AβO solutions demonstrated that while Aβ is essential for memory loss, the fibrillar Aβ in amyloid deposits is not the agent. The AD-like cellular pathologies induced by AβOs suggest their impact provides a unifying mechanism for AD pathogenesis, explaining why early stage disease is specific for memory and accounting for major facets of AD neuropathology. Alternative ideas for triggering mechanisms are being actively investigated. Some research favors insertion of AβOs into membrane, while other evidence supports ligand-like accumulation at particular synapses. Over a dozen candidate toxin receptors have been proposed. AβO binding triggers a redistribution of critical synaptic proteins and induces hyperactivity in metabotropic and ionotropic glutamate receptors. This leads to Ca2+ overload and instigates major facets of AD neuropathology, including tau hyperphosphorylation, insulin resistance, oxidative stress, and synapse loss. Because different species of AβOs have been identified, a remaining question is which oligomer is the major pathogenic culprit. The possibility has been raised that more than one species plays a role. Despite some key unknowns, the clinical relevance of AβOs has been established, and new studies are beginning to point to co-morbidities such as diabetes and hypercholesterolemia as etiological factors. Because pathogenic AβOs appear early in the disease, they offer appealing targets for therapeutics and diagnostics. Promising therapeutic strategies include use of CNS insulin signaling enhancers to protect against the presence of toxins and elimination of the toxins through use of highly specific AβO antibodies. An AD-dependent accumulation of AβOs in CSF suggests their potential use as biomarkers and new AβO probes are opening the door to brain imaging. Overall, current evidence indicates that Aβ oligomers provide a substantive molecular basis for the cause, treatment and diagnosis of Alzheimer’s disease.

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“…In addition, the utility of SMPL Nanodiscs for probing the functional characteristics of membrane proteins in heterogeneous populations is demonstrated by their recent use in incorporating membrane proteins from synapses, used for mechanistic studies of Aβ oligomer receptors and drug discovery ([35], manuscript in preparation).…”

Section: Resultsmentioning

confidence: 99%

Nanodisc-solubilized membrane protein library reflects the membrane proteome

et al. 2013

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The isolation and identification of unknown membrane proteins offers the prospect of discovering new pharmaceutical targets and identifying key biochemical receptors. However, interactions between membrane protein targets and soluble ligands are difficult to study in vitro due to the insolubility of membrane proteins in non-detergent systems. Nanodiscs, nanoscale discoidal lipid bilayers encircled by a membrane scaffold protein belt, have proven to be an effective platform to solubilize membrane proteins and have been used to study a wide variety of purified membrane proteins. This report details the incorporation of an unbiased population of membrane proteins from Escherichia coli membranes into Nanodiscs. This solubilized membrane protein library (SMPL) forms a soluble in vitro model of the membrane proteome. Since Nanodiscs contain isolated proteins or small complexes, the SMPL is an ideal platform for interactomics studies and pull-down assays of membrane proteins. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the protein population before and after formation of the Nanodisc library indicates that a large percentage of the proteins are incorporated into the library. Proteomic identification of several prominent bands demonstrates the successful incorporation of outer and inner membrane proteins into the Nanodisc library.

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Nanoscale Synaptic Membrane Mimetic Allows Unbiased High Throughput Screen That Targets Binding Sites for Alzheimer’s-Associated Aβ Oligomers

Cited by 30 publications

References 68 publications

Nanodiscs in Membrane Biochemistry and Biophysics

Nanodiscs in Membrane Biochemistry and Biophysics

Amyloid β oligomers in Alzheimer’s disease pathogenesis, treatment, and diagnosis

Nanodisc-solubilized membrane protein library reflects the membrane proteome

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