Protocol to identify drug-binding sites in proteins using solution NMR spectroscopy

Dusquetide is a next-generation IDR (innate defense regulator) targeting the major autophagy receptor protein SQSTM1/p62 and modulating the innate immune response. Here, we describe a protocol for determining dusquetide-binding sites of p62 by solution NMR spectroscopy. Step-by-step technique detail...

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Detalles Bibliográficos
Autores principales: Penumutchu, Srinivasa, Liu, Jiuyang, Singh, Upendra K., Kutateladze, Tatiana G., Zhang, Yi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9667315/
https://www.ncbi.nlm.nih.gov/pubmed/36595882
http://dx.doi.org/10.1016/j.xpro.2022.101842
Descripción
Sumario:Dusquetide is a next-generation IDR (innate defense regulator) targeting the major autophagy receptor protein SQSTM1/p62 and modulating the innate immune response. Here, we describe a protocol for determining dusquetide-binding sites of p62 by solution NMR spectroscopy. Step-by-step technique details were provided, including sample preparation, NMR experiment setup, data processing, and binding site analysis. This protocol could be applied to characterize other small molecules targeting the ZZ domain of p62 (9 kDa) or other proteins containing ZZ domains. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2022).

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