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The Use of Co-immunoprecipitation to Study Conformation-Specific Protein Interactions of Oligomeric α-Synuclein Aggregates

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Co-Immunoprecipitation Methods for Brain Tissue

Part of the book series: Neuromethods ((NM,volume 144))

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Abstract

The development of aggregates of specific disease-associated proteins represents a common denominator for many neurodegenerative disorders. The gain of function of the aggregates is hypothesized to initiate pro-degenerative signaling pathways that cause neuronal dysfunctions and ultimately death of affected neurons. Comparing the protein interactome of the native normal functioning disease-associated protein to the interactome of the aggregated forms of the same protein may reveal disease-conducting signaling hubs of relevance to specific diseases. Here, we describe the experimental setup we used to identify specific interaction partners of soluble oligomeric α-synuclein aggregates including step-by-step protocols for preparation of antibody-conjugated Sepharose beads, purification of recombinant soluble α-synuclein oligomers, preparation of synaptosomal extracts from porcine brain, and the actual co-immunoprecipitation. Our goal is to present the reader issues for consideration before starting co-immunoprecipitation experiments and a practical overview of the technical finesses. This approach can be applied to study interaction of any purified disease-linked soluble aggregates.

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Author information

Authors and Affiliations

  1. Danish Research Institute of Translational Neuroscience—DANDRITE, Aarhus University, Aarhus, Denmark

    Cristine Betzer, Rikke Hahn Kofoed & Poul Henning Jensen

  2. Department of Biomedicine, Aarhus University, Aarhus, Denmark

    Cristine Betzer, Rikke Hahn Kofoed & Poul Henning Jensen

Authors
  1. Cristine Betzer
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  2. Rikke Hahn Kofoed
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  3. Poul Henning Jensen
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Corresponding authors

Correspondence to Cristine Betzer or Poul Henning Jensen .

Editor information

Editors and Affiliations

  1. Department of Psychiatry, Saitama Medical University, Saitama, Japan

    Yuji Odagaki

  2. Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden

    Dasiel O. Borroto-Escuela

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Betzer, C., Kofoed, R.H., Jensen, P.H. (2019). The Use of Co-immunoprecipitation to Study Conformation-Specific Protein Interactions of Oligomeric α-Synuclein Aggregates. In: Odagaki, Y., Borroto-Escuela, D. (eds) Co-Immunoprecipitation Methods for Brain Tissue . Neuromethods, vol 144. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8985-0_3

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  • DOI: https://doi.org/10.1007/978-1-4939-8985-0_3

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  • Print ISBN: 978-1-4939-8984-3

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