Proteomics analysis of immuno-precipitated synaptic protein complexes

• Published in: Journal of proteomics 2009-02, Vol.72 (1), p.82-90
• Main Authors: Klemmer, P., Smit, A.B., Li, K.W.
• Format: Article
• Language: English
• Subjects: Animals; Brain; Calcium; Calcium channels (voltage-gated); Cerebral Cortex - metabolism; Glutamic acid receptors; Immuno-precipitation; Immunoprecipitation; Integration; KCNQ2 protein; MALDI tof/tof MS; Mice; Multiprotein Complexes - metabolism; Nerve Tissue Proteins - metabolism; Nerves; Neurotransmission; Organelles; Plasticity (synaptic); Postsynaptic density proteins; Potassium; Potassium channels (voltage-gated); Protein Binding; Protein complexes; Protein interaction; Proteome - metabolism; Proteomics; Synapse; Synapses; Synapses - metabolism; Synaptic vesicles; Synaptic Vesicles - metabolism; Synaptophysin; Voltage
• ISSN: 1874-3919
• DOI: 10.1016/j.jprot.2008.10.005

Synapses are key neuronal elements of the brain. They are responsible for transmission, integration, and storage of information between nerve cells. A synapse is considered as the most complex cellular organelle consisting of ~ 1500 of proteins that are interacting in an activity dependent manner. We have initiated a series of immuno-precipitation experiments in conjunction with LC-MS/MS analysis in order to gain better insight into the organization of the synapse. In particular, we focused on proteins that have been implicated previously in the process of neuroplasticity, i.e., the glutamate receptor (GluR2), scaffolding proteins (PSD-95 and CASK), voltage gated potassium (KCNQ2 and Kv4.2) and calcium (CaV β4) channel subunits, the signalling protein (GIT1) and synaptic vesicle protein (synaptophysin). This study confirms the previous reported protein–protein interactions and furthermore detects novel interactors. In conjunction with the literature reported protein–protein interaction a simple synaptic protein interactome was constructed. This model implicates the potential interaction of distinct protein complexes, and the engagement of single proteins, especially the scaffolding proteins, in multiple protein complexes.