Channel Proteomes

Project

Cav2 Nano-Environments

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

Project Overview

Name Cav2 Nano-Environments
Title

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

Publication

http://dx.doi.org/10.1073/pnas.1005940107

Description

Local Ca2+ signaling occurring within nanometers of voltage-gated Ca2+ (Cav) channels is crucial for CNS function, yet the molecular composition of Cav channel nano-environments is largely unresolved. Here we employed a novel proteomic strategy combining knockout-controlled multi-epitope affinity purifications with high-resolution quantitative mass spectrometry 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 pore-forming alpha-1 and auxiliary beta 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. 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 Ca2+-signaling by Cav2 channels in the brain.

Target Proteins

Protein Name Uniprot/SwissProt
Cav2.1 Voltage-dependent P/Q-type calcium channel subunit alpha-1A P54282
Cav2.2 Voltage-dependent N-type calcium channel subunit alpha-1B Q02294
Cav2.3 Voltage-dependent R-type calcium channel subunit alpha-1E Q07652
Cav beta-1 Voltage-dependent L-type calcium channel subunit beta-1 P54283
Cav beta-2 Voltage-dependent L-type calcium channel subunit beta-2 Q8VGC3
Cav beta-3 Voltage-dependent L-type calcium channel subunit beta-3 P54287
Cav beta-4 Voltage-dependent L-type calcium channel subunit beta-4 Q8R0S4
View all proteins related to this project

Antibodies

Name Supplier
Anti-Cav2.1-a J. Striessnig, Universität Innsbruck, Austria
Anti-Cav2.1-b Millipore (Chemicon), USA
Anti-Cav2.1-c H.G. Knaus, Medizinische Universität Innsbruck, Austria
Anti-Cav2.1-d H.G. Knaus, Medizinische Universität Innsbruck, Austria
Anti-Cav2.2-a Millipore (Chemicon), USA
Anti-Cav2.2-b H.G. Knaus, Medizinische Universität Innsbruck, Austria
Anti-Cav2.2-c H.G. Knaus, Medizinische Universität Innsbruck, Austria
Anti-Cav2.3-a Santa Cruz Biotechnology, USA
Anti-Cav2.3-b Alomone Labs, Israel
Anti-Cav2.x Abcam, USA
Anti-CavBeta1 V. Flockerzi, Universität des Saarlands, Germany
Anti-CavBeta2 V. Flockerzi, Universität des Saarlands, Germany
Anti-CavBeta3 V. Flockerzi, Universität des Saarlands, Germany
Anti-CavBeta4 V. Flockerzi, Universität des Saarlands, Germany
View detailed antibody list

Solubilization Buffers

Name Supplier
ComplexioLyte 114 Logopharm GmbH, Germany
ComplexioLyte 91 Logopharm GmbH, Germany
Digitonin Biosynth AG, Switzerland