Potentiating tweezer affinity to a protein interface with sequence-defined macromolecules on nanoparticles

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Item Type:	Article
Title:	Potentiating tweezer affinity to a protein interface with sequence-defined macromolecules on nanoparticles
Creators Name:	Seiler, T., Lennartz, A., Klein, K., Hommel, K., Figueroa Bietti, A., Hadrovic, I., Kollenda, S., Sager, J., Beuck, C., Chlosta, E., Bayer, P., Juul-Madsen, K., Vorup-Jensen, T., Schrader, T., Epple, M., Knauer, S.K. and Hartmann, L.
Abstract:	Survivin, a well-known member of the inhibitor of apoptosis protein family, is upregulated in many cancer cells, which is associated with resistance to chemotherapy. To circumvent this, inhibitors are currently being developed to interfere with the nuclear export of survivin by targeting its protein-protein interaction (PPI) with the export receptor CRM1. Here, we combine for the first time a supramolecular tweezer motif, sequence-defined macromolecular scaffolds, and ultrasmall Au nanoparticles (us-AuNPs) to tailor a high avidity inhibitor targeting the survivin-CRM1 interaction. A series of biophysical and biochemical experiments, including surface plasmon resonance measurements and their multivalent evaluation by EVILFIT, reveal that for divalent macromolecular constructs with increasing linker distance, the longest linkers show superior affinity, slower dissociation, as well as more efficient PPI inhibition. As a drawback, these macromolecular tweezer conjugates do not enter cells, a critical feature for potential applications. The problem is solved by immobilizing the tweezer conjugates onto us-AuNPs, which enables efficient transport into HeLa cells. On the nanoparticles, the tweezer valency rises from 2 to 16 and produces a 100-fold avidity increase. The hierarchical combination of different scaffolds and controlled multivalent presentation of supramolecular binders was the key to the development of highly efficient survivin-CRM1 competitors. This concept may also be useful for other PPIs.
Keywords:	Cell Nucleus, Cell Nucleus Active Transport, Gold, HeLa Cells, Inhibitor of Apoptosis Proteins, Macromolecular Substances, Metal Nanoparticles, Survivin
Source:	Biomacromolecules
ISSN:	1526-4602
Publisher:	American Chemical Society
Volume:	24
Number:	8
Page Range:	3666-3679
Date:	14 August 2023
Official Publication:	https://doi.org/10.1021/acs.biomac.3c00393
PubMed:	View item in PubMed

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