Mass Spectrometry of RNA-Binding Proteins during Liquid-Liquid Phase Separation Reveals Distinct Assembly Mechanisms and Droplet Architectures

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt

Standard

Mass Spectrometry of RNA-Binding Proteins during Liquid-Liquid Phase Separation Reveals Distinct Assembly Mechanisms and Droplet Architectures. / Sahin, Cagla; Motso, Aikaterini; Gu, Xinyu; Feyrer, Hannes; Lama, Dilraj; Arndt, Tina; Rising, Anna; Gese, Genis Valentin; Haellberg, B. Martin; Marklund, Erik. G.; Schafer, Nicholas P.; Petzold, Katja; Teilum, Kaare; Wolynes, Peter G.; Landreh, Michael.

I: Journal of the American Chemical Society, Bind 145, Nr. 19, 2023.

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt

Harvard

Sahin, C, Motso, A, Gu, X, Feyrer, H, Lama, D, Arndt, T, Rising, A, Gese, GV, Haellberg, BM, Marklund, EG, Schafer, NP, Petzold, K, Teilum, K, Wolynes, PG & Landreh, M 2023, 'Mass Spectrometry of RNA-Binding Proteins during Liquid-Liquid Phase Separation Reveals Distinct Assembly Mechanisms and Droplet Architectures', Journal of the American Chemical Society, bind 145, nr. 19. https://doi.org/10.1021/jacs.3c00932

APA

Sahin, C., Motso, A., Gu, X., Feyrer, H., Lama, D., Arndt, T., Rising, A., Gese, G. V., Haellberg, B. M., Marklund, E. G., Schafer, N. P., Petzold, K., Teilum, K., Wolynes, P. G., & Landreh, M. (2023). Mass Spectrometry of RNA-Binding Proteins during Liquid-Liquid Phase Separation Reveals Distinct Assembly Mechanisms and Droplet Architectures. Journal of the American Chemical Society, 145(19). https://doi.org/10.1021/jacs.3c00932

Vancouver

Sahin C, Motso A, Gu X, Feyrer H, Lama D, Arndt T o.a. Mass Spectrometry of RNA-Binding Proteins during Liquid-Liquid Phase Separation Reveals Distinct Assembly Mechanisms and Droplet Architectures. Journal of the American Chemical Society. 2023;145(19). https://doi.org/10.1021/jacs.3c00932

Author

Sahin, Cagla ; Motso, Aikaterini ; Gu, Xinyu ; Feyrer, Hannes ; Lama, Dilraj ; Arndt, Tina ; Rising, Anna ; Gese, Genis Valentin ; Haellberg, B. Martin ; Marklund, Erik. G. ; Schafer, Nicholas P. ; Petzold, Katja ; Teilum, Kaare ; Wolynes, Peter G. ; Landreh, Michael. / Mass Spectrometry of RNA-Binding Proteins during Liquid-Liquid Phase Separation Reveals Distinct Assembly Mechanisms and Droplet Architectures. I: Journal of the American Chemical Society. 2023 ; Bind 145, Nr. 19.

Bibtex

@article{67d26ba529aa419cb6ffd870bae523a3,
title = "Mass Spectrometry of RNA-Binding Proteins during Liquid-Liquid Phase Separation Reveals Distinct Assembly Mechanisms and Droplet Architectures",
abstract = "Liquid-liquid phase separation (LLPS) of hetero-geneous ribonucleoproteins (hnRNPs) drives the formation of membraneless organelles, but structural information about their assembled states is still lacking. Here, we address this challenge through a combination of protein engineering, native ion mobility mass spectrometry, and molecular dynamics simulations. We used an LLPS-compatible spider silk domain and pH changes to control the self-assembly of the hnRNPs FUS, TDP-43, and hCPEB3, which are implicated in neurodegeneration, cancer, and memory storage. By releasing the proteins inside the mass spectrometer from their native assemblies, we could monitor conformational changes associated with liquid-liquid phase separation. We find that FUS monomers undergo an unfolded-to-globular transition, whereas TDP-43 oligomerizes into partially disordered dimers and trimers. hCPEB3, on the other hand, remains fully disordered with a preference for fibrillar aggregation over LLPS. The divergent assembly mechanisms revealed by ion mobility mass spectrometry of soluble protein species that exist under LLPS conditions suggest structurally distinct complexes inside liquid droplets that may impact RNA processing and translation depending on biological context.",
keywords = "PRION-LIKE DOMAINS, TDP-43, FIBRILS",
author = "Cagla Sahin and Aikaterini Motso and Xinyu Gu and Hannes Feyrer and Dilraj Lama and Tina Arndt and Anna Rising and Gese, {Genis Valentin} and Haellberg, {B. Martin} and Marklund, {Erik. G.} and Schafer, {Nicholas P.} and Katja Petzold and Kaare Teilum and Wolynes, {Peter G.} and Michael Landreh",
year = "2023",
doi = "10.1021/jacs.3c00932",
language = "English",
volume = "145",
journal = "Journal of the American Chemical Society",
issn = "0002-7863",
publisher = "ACS Publications",
number = "19",

}

RIS

TY - JOUR

T1 - Mass Spectrometry of RNA-Binding Proteins during Liquid-Liquid Phase Separation Reveals Distinct Assembly Mechanisms and Droplet Architectures

AU - Sahin, Cagla

AU - Motso, Aikaterini

AU - Gu, Xinyu

AU - Feyrer, Hannes

AU - Lama, Dilraj

AU - Arndt, Tina

AU - Rising, Anna

AU - Gese, Genis Valentin

AU - Haellberg, B. Martin

AU - Marklund, Erik. G.

AU - Schafer, Nicholas P.

AU - Petzold, Katja

AU - Teilum, Kaare

AU - Wolynes, Peter G.

AU - Landreh, Michael

PY - 2023

Y1 - 2023

N2 - Liquid-liquid phase separation (LLPS) of hetero-geneous ribonucleoproteins (hnRNPs) drives the formation of membraneless organelles, but structural information about their assembled states is still lacking. Here, we address this challenge through a combination of protein engineering, native ion mobility mass spectrometry, and molecular dynamics simulations. We used an LLPS-compatible spider silk domain and pH changes to control the self-assembly of the hnRNPs FUS, TDP-43, and hCPEB3, which are implicated in neurodegeneration, cancer, and memory storage. By releasing the proteins inside the mass spectrometer from their native assemblies, we could monitor conformational changes associated with liquid-liquid phase separation. We find that FUS monomers undergo an unfolded-to-globular transition, whereas TDP-43 oligomerizes into partially disordered dimers and trimers. hCPEB3, on the other hand, remains fully disordered with a preference for fibrillar aggregation over LLPS. The divergent assembly mechanisms revealed by ion mobility mass spectrometry of soluble protein species that exist under LLPS conditions suggest structurally distinct complexes inside liquid droplets that may impact RNA processing and translation depending on biological context.

AB - Liquid-liquid phase separation (LLPS) of hetero-geneous ribonucleoproteins (hnRNPs) drives the formation of membraneless organelles, but structural information about their assembled states is still lacking. Here, we address this challenge through a combination of protein engineering, native ion mobility mass spectrometry, and molecular dynamics simulations. We used an LLPS-compatible spider silk domain and pH changes to control the self-assembly of the hnRNPs FUS, TDP-43, and hCPEB3, which are implicated in neurodegeneration, cancer, and memory storage. By releasing the proteins inside the mass spectrometer from their native assemblies, we could monitor conformational changes associated with liquid-liquid phase separation. We find that FUS monomers undergo an unfolded-to-globular transition, whereas TDP-43 oligomerizes into partially disordered dimers and trimers. hCPEB3, on the other hand, remains fully disordered with a preference for fibrillar aggregation over LLPS. The divergent assembly mechanisms revealed by ion mobility mass spectrometry of soluble protein species that exist under LLPS conditions suggest structurally distinct complexes inside liquid droplets that may impact RNA processing and translation depending on biological context.

KW - PRION-LIKE DOMAINS

KW - TDP-43

KW - FIBRILS

U2 - 10.1021/jacs.3c00932

DO - 10.1021/jacs.3c00932

M3 - Journal article

C2 - 37145883

VL - 145

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

SN - 0002-7863

IS - 19

ER -

ID: 347477935