Expression, purification and structural analysis of functional GABA transporter 1 using the baculovirus expression system

• Jing Hu,
• Chris Weise,
• Christoph Böttcher,
• Hua Fan and
• Jian Yin

Beilstein J. Org. Chem. 2017, 13, 874–882, doi:10.3762/bjoc.13.88

• (GFP) fusion protein was functionally expressed in insect Sf9 cells by the BAC-TO-BAC® baculovirus expression system. A two-step procedure to purify the GAT1/GFP fusion protein from insect Sf9 cells has been established and involves immunoaffinity chromatography using self-prepared anti-GFP antibodies

• isolated in its monomeric form. Keywords: baculovirus expression system; chromatography; γ-aminobutyric acid (GABA); GABA transporter 1 (GAT1); Introduction γ-Aminobutyric acid (GABA) is the most abundant inhibitory neurotransmitter in the central nervous system (CNS) of vertebrate species. GABAergic

• proteins should be in a native and functional conformation during expression and purification. The natural abundance of most membrane proteins is typically not high enough for the isolation of sufficient quantities for functional and structural studies. In this work, the baculovirus expression system was