TY - JOUR
T1 - Disentangling the recognition complexity of a protein hub using a nanopore
AU - Mayse, Lauren Ashley
AU - Imran, Ali
AU - Larimi, Motahareh Ghahari
AU - Cosgrove, Michael S.
AU - Wolfe, Aaron James
AU - Movileanu, Liviu
N1 - Funding Information:
We thank our colleagues in the Movileanu and Cosgrove laboratories and at Ichor Life Sciences laboratories for their comments on the manuscript and stimulating discussions as well as for their technical assistance during the very early stage of this project. This work was supported by the National Cancer Institute of the U.S. National Institutes of Health grant R01 CA140522 (to M.S.C) and by the National Institute of General Medical Sciences of the U.S. National Institutes of Health grant R01 GM129429 (to L.M.).
Publisher Copyright:
© 2022, The Author(s).
PY - 2022/12
Y1 - 2022/12
N2 - 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 (MLL4Win), a WDR5 ligand. Our approach reveals a broad dynamic range of MLL4Win-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.
AB - 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 (MLL4Win), a WDR5 ligand. Our approach reveals a broad dynamic range of MLL4Win-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.
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U2 - 10.1038/s41467-022-28465-8
DO - 10.1038/s41467-022-28465-8
M3 - Article
C2 - 35190547
AN - SCOPUS:85125156028
VL - 13
JO - Nature Communications
JF - Nature Communications
SN - 2041-1723
IS - 1
M1 - 978
ER -