Targeted tandem affinity purification of PSD-95 recovers core postsynaptic complexes and schizophrenia susceptibility proteins (original) (raw)
Fernandez, E., Collins, M.O. orcid.org/0000-0002-7656-4975, Uren, R.T. et al. (7 more authors) (2009)Targeted tandem affinity purification of PSD-95 recovers core postsynaptic complexes and schizophrenia susceptibility proteins. Molecular Systems Biology, 5 (1). 269.
Abstract
The molecular complexity of mammalian proteomes demands new methods for mapping the organization of multiprotein complexes. Here, we combine mouse genetics and proteomics to characterize synapse protein complexes and interaction networks. New tandem affinity purification (TAP) tags were fused to the carboxyl terminus of PSD‐95 using gene targeting in mice. Homozygous mice showed no detectable abnormalities in PSD‐95 expression, subcellular localization or synaptic electrophysiological function. Analysis of multiprotein complexes purified under native conditions by mass spectrometry defined known and new interactors: 118 proteins comprising crucial functional components of synapses, including glutamate receptors, K+ channels, scaffolding and signaling proteins, were recovered. Network clustering of protein interactions generated five connected clusters, with two clusters containing all the major ionotropic glutamate receptors and one cluster with voltage‐dependent K+ channels. Annotation of clusters with human disease associations revealed that multiple disorders map to the network, with a significant correlation of schizophrenia within the glutamate receptor clusters. This targeted TAP tagging strategy is generally applicable to mammalian proteomics and systems biology approaches to disease.
Metadata
| Authors/Creators: | Fernandez, E.Collins, M.O.  https://orcid.org/0000-0002-7656-4975Uren, R.T.Kopanitsa, M.V.Komiyama, N.H.Croning, M.D.R.Zografos, L.Armstrong, J.D.Choudhary, J.S.Grant, S.G.N. |
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| Copyright, Publisher and Additional Information: | © 2009 EMBO and Macmillan Publishers Limited. This article is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 Licence (http://creativecommons.org/licenses/by-nc-nd/3.0/). |
| Keywords: | gene targeting; postsynaptic complexes; postsynaptic density-95; schizophrenia; tandem affinity purification |
| Dates: | Accepted: 1 April 2009Published: 19 May 2009 |
| Institution: | The University of Sheffield |
| Academic Units: | The University of Sheffield > Faculty of Science (Sheffield) > School of Biosciences (Sheffield) > Department of Biomedical Science (Sheffield) |
| Depositing User: | Symplectic Sheffield |
| Date Deposited: | 10 Feb 2020 11:43 |
| Last Modified: | 11 Feb 2020 04:50 |
| Status: | Published |
| Publisher: | EMBO Press |
| Refereed: | Yes |
| Identification Number: | https://doi.org/10.1038/msb.2009.27 |
| Related URLs: | Author |