LENDS, Alons; DASKALOV, Asen; MALECKIS, Ansis; DELAMARE, Aline; BERBON, Mélanie; GRELARD, Axelle; MORVAN, Estelle; SHENOY, Jayakrishna; DUTOUR, Antoine; TOLCHARD, James; NOUBHANI, Abdelmajid; GIRAUD, Marie-France; SANCHEZ, Corinne; HABENSTEIN, Birgit; GUICHARD, Gilles; COMPAIN, Guillaume; JAUDZEMS, Kristaps; SAUPE, Sven J.; LOQUET, Antoine

doi:10.1038/s42003-022-04175-1

LENDS, Alons
Chimie et Biologie des Membranes et des Nanoobjets [CBMN]

DASKALOV, Asen

Chimie et Biologie des Membranes et des Nanoobjets [CBMN]

MALECKIS, Ansis

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Article de revue

Ce document a été publié dans

Communications Biology. 2022-11-09, vol. 5, n° 1, p. 1-10

Résumé en anglais

Structural investigations of amyloid fibrils often rely on heterologous bacterial overexpression of the protein of interest. Due to their inherent hydrophobicity and tendency to aggregate as inclusion bodies, many amyloid proteins are challenging to express in bacterial systems. Cell-free protein expression is a promising alternative to classical bacterial expression to produce hydrophobic proteins and introduce NMR-active isotopes that can improve and speed up the NMR analysis. Here we implement the cell-free synthesis of the functional amyloid prion HET-s(218-289). We present an interesting case where HET-s(218-289) directly assembles into infectious fibril in the cell-free expression mixture without the requirement of denaturation procedures and purification. By introducing tailored 13C and 15N isotopes or CF3 and 13CH2F labels at strategic amino-acid positions, we demonstrate that cell-free synthesized amyloid fibrils are readily amenable to high-resolution magic-angle spinning NMR at sub-milligram quantity.< Réduire

URI

https://oskar-bordeaux.fr/handle/20.500.12278/172642

DOI

http://dx.doi.org/10.1038/s42003-022-04175-1

Projet Européen

European Union (ERDF, INTERREGV)

Project ANR

Structure et fonctions de motifs amyloïdes impliqués dans la transduction du signal
Interactions supramoléculaires sélectives et directionnelles à base de synthons fluorés hautement polaires

Unités de recherche

CBMN : Chimie & de Biologie des Membranes & des Nano-objets - UMR 5248

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Cell-free synthesis of amyloid fibrils with infectious properties and amenable to sub-milligram magic-angle spinning NMR analysis