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.
Publication types
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Evaluation Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Antigens, CD / chemistry
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Antigens, CD / genetics
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Antigens, CD / metabolism*
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Cell Line
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GPI-Linked Proteins / chemistry
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GPI-Linked Proteins / genetics
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GPI-Linked Proteins / metabolism
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HIV Infections / genetics
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HIV Infections / metabolism*
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HIV Infections / virology
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HIV-1 / genetics
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HIV-1 / metabolism*
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High-Throughput Screening Assays / methods*
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Human Immunodeficiency Virus Proteins / genetics
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Human Immunodeficiency Virus Proteins / metabolism*
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Humans
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Protein Binding
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Protein Structure, Tertiary
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Viral Regulatory and Accessory Proteins / genetics
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Viral Regulatory and Accessory Proteins / metabolism*
Substances
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Antigens, CD
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BST2 protein, human
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GPI-Linked Proteins
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Human Immunodeficiency Virus Proteins
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Viral Regulatory and Accessory Proteins
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vpu protein, Human immunodeficiency virus 1