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Identification and Characterization of Protein Complexes from Total Brain and Synaptoneurosomes: Heterogeneity of Molecular Complexes in Distinct Subcellular Domains

  • Silvia De Rubeis
  • Claudia BagniEmail author
Protocol
Part of the Neuromethods book series (NM, volume 57)

Abstract

Neurons are highly polarized cells characterized by subcellular microdomains: the synapses. These compartments are specialized structures and are, for certain cellular pathways, independent from the cell body. To achieve such a functional specificity, including local mRNA translation, different molecular complexes are transported along the dendrites and locally regulated. Characterization of such a molecular diversity may help to elucidate neuronal functions as well as detect differences in neuronal dysfunctions. Here, we describe a method to specifically dissect a molecular complex according to the neuronal subcellular compartment. Specifically, the complexes are isolated by immunoprecipitation of the protein of interest from brain lysates or from purified synapses (synaptoneurosomes) and identified by mass spectrometry analysis.

Key words

Immunoprecipitation Synaptoneurosomes Synaptic proteomic Neurites mRNP transport 

Notes

Acknowledgments

This study was supported by Telethon, Compagnia di San Paolo, COFIN and FWO.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Center for Human GeneticsKatholieke Universiteit LeuvenLeuvenBelgium
  2. 2.Department of Molecular and Developmental GeneticsLeuvenBelgium
  3. 3.Department of Experimental Medicine and Biochemical SciencesUniversity of Rome “Tor Vergata”RomeItaly

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