Hydrogen bond guidance and aromatic stacking drive liquid-liquid phase separation of intrinsically disordered histidine-rich peptides

Research output: Contribution to journalArticleScientificpeer-review


  • Bartosz Gabryelczyk
  • Hao Cai
  • Xiangyan Shi
  • Yue Sun
  • Piet J.M. Swinkels
  • Stefan Salentinig
  • Konstantin Pervushin
  • Ali Miserez

Research units

  • Wageningen University & Research
  • Swiss Federal Laboratories for Materials Science and Technology (Empa)
  • University of Fribourg
  • Nanyang Technological University


Liquid-liquid phase separation (LLPS) of intrinsically disordered proteins (IDPs) is involved in both intracellular membraneless organelles and extracellular tissues. Despite growing understanding of LLPS, molecular-level mechanisms behind this process are still not fully established. Here, we use histidine-rich squid beak proteins (HBPs) as model IDPs to shed light on molecular interactions governing LLPS. We show that LLPS of HBPs is mediated though specific modular repeats. The morphology of separated phases (liquid-like versus hydrogels) correlates with the repeats’ hydrophobicity. Solution-state NMR indicates that LLPS is a multistep process initiated by deprotonation of histidine residues, followed by transient hydrogen bonding with tyrosine, and eventually by hydrophobic interactions. The microdroplets are stabilized by aromatic clustering of tyrosine residues exhibiting restricted molecular mobility in the nano-to-microsecond timescale according to solid-state NMR experiments. Our findings provide guidelines to rationally design pH-responsive peptides with LLPS ability for various applications, including bioinspired protocells and smart drug-delivery systems.


Original languageEnglish
Article number5465
JournalNature Communications
Issue number1
Publication statusPublished - 1 Dec 2019
MoE publication typeA1 Journal article-refereed

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