Establishment of a high-throughput screening assay for interaction inhibitor between BST-2 and Vpu.
- Author:
Xiao-Jing PANG
1
;
Si-Qi HU
;
Yue ZHANG
;
Shan CEN
;
Qi JIN
;
Fei GUO
Author Information
1. Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, China.
- Publication Type:Journal Article
- MeSH:
Antigens, CD;
chemistry;
genetics;
metabolism;
Cell Line;
GPI-Linked Proteins;
chemistry;
genetics;
metabolism;
HIV Infections;
genetics;
metabolism;
virology;
HIV-1;
genetics;
metabolism;
High-Throughput Screening Assays;
methods;
Human Immunodeficiency Virus Proteins;
genetics;
metabolism;
Humans;
Protein Binding;
Protein Structure, Tertiary;
Viral Regulatory and Accessory Proteins;
genetics;
metabolism
- From:
Chinese Journal of Virology
2012;28(6):633-638
- CountryChina
- Language:Chinese
-
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.
