1. [Establishment of a high-throughput screening assay for interaction inhibitor between BST-2 and Vpu].
- Author
-
Pang XJ, Hu SQ, Zhang Y, Cen S, Jin Q, and Guo F
- Subjects
- Antigens, CD chemistry, Antigens, CD genetics, Cell Line, GPI-Linked Proteins chemistry, GPI-Linked Proteins genetics, GPI-Linked Proteins metabolism, HIV Infections genetics, HIV Infections virology, HIV-1 genetics, Human Immunodeficiency Virus Proteins genetics, Humans, Protein Binding, Protein Structure, Tertiary, Viral Regulatory and Accessory Proteins genetics, Antigens, CD metabolism, HIV Infections metabolism, HIV-1 metabolism, High-Throughput Screening Assays methods, Human Immunodeficiency Virus Proteins metabolism, Viral Regulatory and Accessory Proteins metabolism
- Abstract
BST-2 plays an important role in host innate immune response via inhibiting the release of HIV-1. HIV-1 accessory protein Vpu can interact with BST-2 through its transmembrane domains, degrade BST-2, and decrease BST-2 that are transported to the cell surface, thus anti-virus function of BST-2 is antagonized. In our study, we constructed plasmid RB connecting Rluc to the N-termimal of BST-2, and plasmid VE connecting EYFP to the C-terminal of Vpu. The two fusion proteins were co-expressed in 293 cells, and the interaction between the two proteins was detected via BRET method. And we further established a stable 293 cell line of dual-expression. By using BRET method, and the interaction between BST-2 and Vpu transmembrane domain as the target, a high-throughput screening assay was created that was expected to seek novel interaction inhibitors.
- Published
- 2012