Juneyoung Jung
Kyung Hee University, South Korea
Title: Bacterial overexpression and purification of Vps34-binding domain of Beclin 1
Biography
Biography: Juneyoung Jung
Abstract
Beclin 1 recruits several autophagy-specific factors such as ATG14L and UVRAG, on the catalytic subunit of Class III Phosphatidylinositol-3-kinase (known as Vps34) to form an autophagy-initiating PIK3C3/VPS34 complex. Beclin 1 contains several characteristic protein-protein interaction domains for a variety of proteins that determine the function of the resulting complex. Therefore, the investigation of these domains is important to understand the roles of Beclin 1 as a scaffolding subunit of PIK3C3/VPS34 complex. In the present study, we have prepared the bacterial overexpression system to obtain Vps34-binding domain of Beclin 1 (Vps34-BD), but the proteins was quite unstable and presented as an inclusion body in E. coli. With several condition tests for protein expression and purification, we have set up the optimized protocol with the denaturing purification method. Vps34-binding assay has confirmed that the bacterially-purified Vps34-BD is soluble and functional.