A novel binding site between the voltage-dependent calcium channel Ca_V1.2 subunit and Ca_Vβ2 subunit discovered using a new analysis method for protein-protein interactions.

ABSTRACT: We developed a new method to analyze protein-protein interactions using a dual-inducible prokaryotic expression system. To evaluate protein-protein binding, a chimeric fusion toxin gene was constructed using a DNase-treated short DNA fragment (epitope library) and CcdB, which encodes a DNA topoisomerase II toxin. Protein-protein interactions would affect toxin activity, resulting in colony formation. Using this novel system, we found a new binding site in the voltage-dependent calcium channel α1 subunit (Ca_V1.2) for the voltage-dependent calcium channel β2 subunit. Prokaryotic expression screening of the β2 subunit using an epitope library of Ca_V1.2 resulted in two overlapping clones of the C-terminal sequence of Ca_V1.2. In vitro overlay and immunoprecipita

SUBMITTER: Murakami AM 

PROVIDER: S-EPMC10460381 | biostudies-literature | 2023 Aug

REPOSITORIES: biostudies-literature