A fluorescence polarization assay for high-throughput screening of inhibitors against HIV-1 Nef-mediated MHC-I downregulation.
J Biol Chem
; 300(8): 107529, 2024 Aug.
Article
en En
| MEDLINE
| ID: mdl-38960039
ABSTRACT
The multifunctional, HIV-1 accessory protein Nef enables infected cells to evade host immunity and thus plays a key role in viral pathogenesis. One prominent function of Nef is the downregulation of major histocompatibility complex class I (MHC-I), which disrupts antigen presentation and thereby allows the infected cells to evade immune surveillance by the cytotoxic T cells. Therapeutic inhibition of this Nef function is a promising direction of antiretroviral drug discovery as it may revitalize cytotoxic T cells to identify, and potentially clear, hidden HIV-1 infections. Guided by the crystal structure of the protein complex formed between Nef, MHC-I, and the hijacked clathrin adaptor protein complex 1, we have developed a fluorescence polarization-based assay for inhibitor screening against Nef's activity on MHC-I. The optimized assay has a good signal-to-noise ratio, substantial tolerance of dimethylsulfoxide, and excellent ability to detect competitive inhibition, indicating that it is suitable for high-throughput screening.
Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Antígenos de Histocompatibilidad Clase I
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Regulación hacia Abajo
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VIH-1
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Productos del Gen nef del Virus de la Inmunodeficiencia Humana
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Ensayos Analíticos de Alto Rendimiento
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Polarización de Fluorescencia
Límite:
Humans
Idioma:
En
Año:
2024
Tipo del documento:
Article