Cucuzza, Stefano, Sitnik, Malgorzata, Jurt, Simon, Michel, Erich, Dai, Wenzhao, Müntener, Thomas, Ernst, Patrick, Häussinger, Daniel, Plückthun, Andreas and Zerbe, Oliver (2023) Unexpected dynamics in femtomolar complexes of binding proteins with peptides. Nature communications, 14 (1). p. 7823. ISSN 2041-1723

Abstract

Ultra-tight binding is usually observed for proteins associating with rigidified molecules. Previously, we demonstrated that femtomolar binders derived from the Armadillo repeat proteins (ArmRPs) can be designed to interact very tightly with fully flexible peptides. Here we show for ArmRPs with four and seven sequence-identical internal repeats that the peptide-ArmRP complexes display conformational dynamics. These dynamics stem from transient breakages of individual protein-residue contacts that are unrelated to overall unbinding. The labile contacts involve electrostatic interactions. We speculate that these dynamics allow attaining very high binding affinities, since they reduce entropic losses. Importantly, only NMR techniques can pick up these local events by directly detecting conformational exchange processes without complications from changes in solvent entropy. Furthermore, we demonstrate that the interaction surface of the repeat protein regularizes upon peptide binding to become more compatible with the peptide geometry. These results provide novel design principles for ultra-tight binders.

Item Type:	Article
Date Deposited:	13 Dec 2023 00:45
Last Modified:	13 Dec 2023 00:45
URI:	https://oak.novartis.com/id/eprint/52604