Identification of Kv4.2 protein complex and modifications by tandem affinity purification-mass spectrometry in primary neurons.

ABSTRACT: Proteins usually form complexes to fulfill variable physiological functions. In neurons, communication relies on synapses where receptors, channels, and anchoring proteins form complexes to precisely control signal transduction, synaptic integration, and action potential firing. Although there are many published protocols to isolate protein complexes in cell lines, isolation in neurons has not been well established. Here we introduce a method that combines lentiviral protein expression with tandem affinity purification followed by mass-spectrometry (TAP-MS) to identify protein complexes in neurons. This protocol can also be used to identify post-translational modifications (PTMs) of synaptic proteins. We used the A-type voltage-gated K⁺ channel subunit Kv4.2 as the target protei

SUBMITTER: Hu JH 

PROVIDER: S-EPMC9788671 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature