Enzymatic encoding of topology in an intrinsically disordered single-chain protein

Research output: Contribution to journal › Research article › Contributed › peer-review

Contributors

  • Joshua Johani - , Leibniz Institute of Polymer Research Dresden, University of Stellenbosch (Author)
  • Kristin Eichelberger - , Chair of Food Engineering (Author)
  • Olga Guskova - , Leibniz Institute of Polymer Research Dresden (Author)
  • Simbulele Charlotte Dunwa - , University of Stellenbosch (Author)
  • Hans Bolinsson - , Lund University (Author)
  • Anja-Maria Börjesdottir - , Lund University (Author)
  • Lars Nilsson - , Lund University (Author)
  • Doris Jaros - , Chair of Food Engineering (Author)
  • Harald Rohm - , Chair of Food Engineering (Author)
  • Albena Lederer - , Leibniz Institute of Polymer Research Dresden, University of Stellenbosch (Author)

Abstract

Controlling the three-dimensional topology of single-chain nanoparticles (SCNPs) remains a central challenge in polymer and protein chemistry, particularly for intrinsically disordered systems lacking defined secondary structure. Here, we demonstrate that selective enzymatic intramolecular cross-linking can encode topologically biased interactions in an intrinsically disordered protein (IDP), yielding compact SCNPs with reproducible cavity architecture. Using β-casein-rich sodium caseinate (βNaCn) as a model surrogate for bovine β-casein (β-Cn), microbial transglutaminase (mTGase) introduces sparse, sequence-resolved glutamine-lysine isopeptide bonds that drive reproducible chain collapse without inducing secondary structure. Analyses by size exclusion chromatography with quintuple detection (SEC-D5), cross-linking mass spectrometry (XL-MS), molecular dynamics (MD) simulations, and SEC coupled to synchrotron small-angle x-ray scattering (SEC-SAXS) converge to reveal a topology combining a stable, compact, hydrophobic core with flexible, disordered loops. These cavities are probed using Nile red (NR) fluorescence and SEC-SAXS, which together provide topology information via guest-induced density redistribution after NR capture. This work establishes that sparse enzymatic constraint installation, combined with residue-resolved cross-link mapping and orthogonal structural analysis, can encode and validate topology in a disordered single chain, thereby placing IDP-like covalent folding in direct conceptual continuity with SCNP design.

Details

Original languageEnglish
Article numbere3738676
Number of pages8
JournalAngewandte Chemie International Edition
Volume65
Issue number28
Early online date26 May 2026
Publication statusPublished - 6 Jul 2026
Peer-reviewedYes

External IDs

Scopus 105040379473
PubMed 42189706

Keywords

ASJC Scopus subject areas

Keywords

  • SEC-SAXS, enzymatic cross-linking, topology encoding, intrinsically disordered proteins, single-chain nanoparticles