9YLZ

Backbone Modification in the Villin Headpiece Miniprotein: HP35 with beta3Leu at Position 28

Summary for 9YLZ

• Entry DOI: 10.2210/pdb9ylz/pdb
• NMR Information: BMRB: 31270
• Descriptor: Villin-1 (1 entity in total)
• Functional Keywords: miniprotein, heterogeneous backbone, proteomimetic, structural protein
• Biological source: Gallus gallus (chicken)
• Total number of polymer chains: 1
• Total formula weight: 4058.70

Authors

Lin, Y.,David, R.M.,Amin, D.M.,Osborne, S.W.J.,Horne, W.S. (deposition date: 2025-10-09, release date: 2026-01-07)

Primary citation

Lin, Y.,David, R.M.,Amin, D.M.,Osborne, S.W.J.,Horne, W.S.
Backbone engineering in the hydrophobic core of villin headpiece.
Rsc Chem Biol, 2025

PubMed Abstract: Changing the backbone connectivity of proteins can impart useful new traits while maintaining essential structural and functional features. In design of artificial proteomimetic agents, backbone modification is usually isolated to sites that are solvent-exposed in the folded state, as similar changes at buried residues can alter the fold. Recent work has shown that core backbone modification without structural perturbation is possible; however, the modifications in that study were consistently destabilizing and made in a prototype of exceptionally high conformational stability. Here, we report efforts to broaden the scope and improve the efficacy of core backbone engineering by applying it to the C-terminal subdomain of villin headpiece. A series of variants are prepared in which different artificial residue types are incorporated at core positions throughout the sequence, including a crucial aromatic triad. Impacts on folding energetics are quantified by biophysical methods, and high-resolution structures of several variants determined by NMR. We go on to construct a variant with ∼40% of its core modified that adopts a fold identical to the prototype while showing enhanced thermodynamic stability.

PubMed: 41450753
DOI: 10.1039/d5cb00269a

Experimental method

SOLUTION NMR