Disentangling the recognition complexity of a protein hub using a nanopore.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 35190547.
- Also identified by DOI 10.1038/s41467-022-28465-8 and PMC identifier 8861093.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
Abstract
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<sub>Win</sub>), a WDR5 ligand. Our approach reveals a broad dynamic range of MLL4<sub>Win</sub>-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.
Medical subject headings
- Intracellular Signaling Peptides and Proteins
- Nanopores
- Protein Engineering