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

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

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

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
Volume10
Issue number1
DOIs
Publication statusPublished - 1 Dec 2019

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histidine
Histidine
Intrinsically Disordered Proteins
Phase separation
Beak
peptides
Hydrogen
Decapodiformes
Hydrogen bonds
liquid phases
hydrogen bonds
Hydrophobic and Hydrophilic Interactions
Peptides
Tyrosine
Liquids
liquids
Artificial Cells
Hydrogels
proteins
Hydrogen Bonding

Cite this

Gabryelczyk, Bartosz ; Cai, Hao ; Shi, Xiangyan ; Sun, Yue ; Swinkels, Piet J.M. ; Salentinig, Stefan ; Pervushin, Konstantin ; Miserez, Ali. / Hydrogen bond guidance and aromatic stacking drive liquid-liquid phase separation of intrinsically disordered histidine-rich peptides. In: Nature Communications. 2019 ; Vol. 10, No. 1.
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abstract = "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.",
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Hydrogen bond guidance and aromatic stacking drive liquid-liquid phase separation of intrinsically disordered histidine-rich peptides. / Gabryelczyk, Bartosz; Cai, Hao; Shi, Xiangyan; Sun, Yue; Swinkels, Piet J.M.; Salentinig, Stefan; Pervushin, Konstantin; Miserez, Ali.

In: Nature Communications, Vol. 10, No. 1, 5465, 01.12.2019.

Research output: Contribution to journalArticleAcademicpeer-review

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T1 - Hydrogen bond guidance and aromatic stacking drive liquid-liquid phase separation of intrinsically disordered histidine-rich peptides

AU - Gabryelczyk, Bartosz

AU - Cai, Hao

AU - Shi, Xiangyan

AU - Sun, Yue

AU - Swinkels, Piet J.M.

AU - Salentinig, Stefan

AU - Pervushin, Konstantin

AU - Miserez, Ali

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AB - 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.

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