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Disentangling the recognition complexity of a protein hub using a nanopore

WD40 repeat proteins are frequently involved in processing cell signaling and scaffolding large multi-subunit machineries. Despite their significance in physiological and disease-like conditions, their reversible interactions with other proteins remain modestly examined. Here, we show the developmen...

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Detalles Bibliográficos
Autores principales: Mayse, Lauren Ashley, Imran, Ali, Larimi, Motahareh Ghahari, Cosgrove, Michael S., Wolfe, Aaron James, Movileanu, Liviu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8861093/
https://www.ncbi.nlm.nih.gov/pubmed/35190547
http://dx.doi.org/10.1038/s41467-022-28465-8
Descripción
Sumario:WD40 repeat proteins are frequently involved in processing cell signaling and scaffolding large multi-subunit machineries. Despite their significance in physiological and disease-like conditions, their reversible interactions with other proteins remain modestly examined. Here, we show the development and validation of a protein nanopore for the detection and quantification of WD40 repeat protein 5 (WDR5), a chromatin-associated hub involved in epigenetic regulation of histone methylation. Our nanopore sensor is equipped with a 14-residue Win motif of mixed lineage leukemia 4 methyltransferase (MLL4(Win)), a WDR5 ligand. Our approach reveals a broad dynamic range of MLL4(Win)-WDR5 interactions and three distant subpopulations of binding events, representing three modes of protein recognition. The three binding events are confirmed as specific interactions using a weakly binding WDR5 derivative and various environmental contexts. These outcomes demonstrate the substantial sensitivity of our nanopore sensor, which can be utilized in protein analytics.