Proteomics analysis of immuno-precipitated synaptic protein complexes

Klemmer, Patricia; Smit, August B.; Li, Ka Wan

doi:10.1016/j.jprot.2008.10.005

Cited by 56 publications

(51 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Sample Preparation for Mass Spectrometry-In-gel digestion procedure, MALDI MS, and protein identification procedure were performed as previously described (23). The peptides were redissolved in 20 l 0.1% TFA and separated on an analytical capillary C18 column (150 mm ϫ 100 m inner diameter column) at 400 nl/min with a linearly increasing concentration of acetonitrile.…”

Section: Mice-sod1mentioning

confidence: 99%

Proteomic Analysis of Dynein-Interacting Proteins in Amyotrophic Lateral Sclerosis Synaptosomes Reveals Alterations in the RNA-Binding Protein Staufen1

Gershoni-Emek

Mazza

Chein

et al. 2016

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

Synapse disruption takes place in many neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS). However, the mechanistic understanding of this process is still limited. We set out to study a possible role for dynein in synapse integrity. Cytoplasmic dynein is a multisubunit intracellular molecule responsible for diverse cellular functions, including long-distance transport of vesicles, organelles, and signaling factors toward the cell center. A less well-characterized role dynein may play is the spatial clustering and anchoring of various factors including mRNAs in distinct cellular domains such as the neuronal synapse. Here, in order to gain insight into dynein functions in synapse integrity and disruption, we performed a screen for novel dynein interactors at the synapse. Dynein immunoprecipitation from synaptic fractions of the ALS model mSOD1 G93A and wild-type controls, followed by mass spectrometry analysis on synaptic fractions of the ALS model mSOD1 G93A and wild-type controls, was performed. Using advanced network analysis, we identified Staufen1, an RNA-binding protein required for the transport and localization of neuronal RNAs, as a major mediator of dynein interactions via its interaction with protein phosphatase 1-beta (PP1B). Both in vitro and in vivo validation assays demonstrate the interactions of Staufen1 and PP1B with dynein, and their colocalization with synaptic markers was altered as a result of two separate ALS-linked mutations: mSOD1 G93A and TDP43 A315T. Taken together, we suggest a model in which dynein's interaction with Staufen1 regulates mRNA localization along the axon and the synapses, and alterations in this process may correlate with synapse disruption and ALS toxicity. Molecular & Cellular

show abstract

Section: Mice-sod1mentioning

confidence: 99%

Proteomic Analysis of Dynein-Interacting Proteins in Amyotrophic Lateral Sclerosis Synaptosomes Reveals Alterations in the RNA-Binding Protein Staufen1

Gershoni-Emek

Mazza

Chein

et al. 2016

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

show abstract

“…Hippocampi were collected and cellular fractionation was performed as previously described (Klemmer et al, 2009). In brief, hippocampi were homogenized in a glass Potter homogenizer containing 5 ml ice-cold homogenization buffer (320 mM sucrose in 5 mM Hepes, pH 7.4) at 900 rpm.…”

Section: Cellular Fractionationmentioning

confidence: 99%

Ubiquitin ligase TRIM3 controls hippocampal plasticity and learning by regulating synaptic γ-actin levels

Schreiber¹,

Végh²,

Dawitz³

et al. 2015

J Exp Med

View full text Add to dashboard Cite

“…Finally, reliable identification of less-abundant or smaller proteins requires high-resolution quantitative MS (24). In the light of these concerns, previous efforts to identify proteins associated with Cav channels (25,26) might have missed important components or contained false-positive interactors.…”

mentioning

confidence: 99%

Quantitative proteomics of the Cav2 channel nano-environments in the mammalian brain

Müller

Haupt

Bildl

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

279

308

View full text Add to dashboard Cite

Local Ca 2+ signaling occurring within nanometers of voltage-gated Ca 2+ (Cav) channels is crucial for CNS function, yet the molecular composition of Cav channel nano-environments is largely unresolved. Here, we used a proteomic strategy combining knockoutcontrolled multiepitope affinity purifications with high-resolution quantitative MS for comprehensive analysis of the molecular nano-environments of the Cav2 channel family in the whole rodent brain. The analysis shows that Cav2 channels, composed of poreforming α1 and auxiliary β subunits, are embedded into protein networks that may be assembled from a pool of ∼200 proteins with distinct abundance, stability of assembly, and preference for the three Cav2 subtypes. The majority of these proteins have not previously been linked to Cav channels; about two-thirds are dedicated to the control of intracellular Ca 2+ concentration, including G proteincoupled receptor-mediated signaling, to activity-dependent cytoskeleton remodeling or Ca 2+ -dependent effector systems that comprise a high portion of the priming and release machinery of synaptic vesicles. The identified protein networks reflect the cellular processes that can be initiated by Cav2 channel activity and define the molecular framework for organization and operation of local Ca 2+ signaling by Cav2 channels in the brain.calcium channel | Ca 2+ signaling | proteome | biochemistry | mass spectrometry

show abstract

Proteomics analysis of immuno-precipitated synaptic protein complexes

Cited by 56 publications

References 43 publications

Proteomic Analysis of Dynein-Interacting Proteins in Amyotrophic Lateral Sclerosis Synaptosomes Reveals Alterations in the RNA-Binding Protein Staufen1

Proteomic Analysis of Dynein-Interacting Proteins in Amyotrophic Lateral Sclerosis Synaptosomes Reveals Alterations in the RNA-Binding Protein Staufen1

Ubiquitin ligase TRIM3 controls hippocampal plasticity and learning by regulating synaptic γ-actin levels

Quantitative proteomics of the Cav2 channel nano-environments in the mammalian brain

Contact Info

Product

Resources

About