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High intracellular stability of the spidroin N-terminal domain in spite of abundant amyloidogenic segments revealed by in-cell hydrogen/deuterium exchange mass spectrometry

Kaldmae, Margit and Leppert, Axel and Chen, Gefei and Sarr, Medoune and Sahin, Cagla and Nordling, Kerstin and Kronqvist, Nina and Gonzalvo-Ulla, Marta and Fritz, Nicolas and Abelein, Axel and Lain, Sonia and Biverstal, Henrik and Jornvall, Hans and Lane, David P. and Rising, Anna and Johansson, Jan and Landreh, Michael (2020). High intracellular stability of the spidroin N-terminal domain in spite of abundant amyloidogenic segments revealed by in-cell hydrogen/deuterium exchange mass spectrometry. FEBS Journal. 287 , 2823-2833
[Journal article]

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Abstract

Proteins require an optimal balance of conformational flexibility and stability in their native environment to ensure their biological functions. A striking example is spidroins, spider silk proteins, which are stored at extremely high concentrations in soluble form, yet undergo amyloid-like aggregation during spinning. Here, we elucidate the stability of the highly soluble N-terminal domain (NT) of major ampullate spidroin 1 in the Escherichia coli cytosol as well as in inclusion bodies containing fibrillar aggregates. Surprisingly, we find that NT, despite being largely composed of amyloidogenic sequences, showed no signs of concentration-dependent aggregation. Using a novel intracellular hydrogen/deuterium exchange mass spectrometry (HDX-MS) approach, we reveal that NT adopts a tight fold in the E. coli cytosol and in this manner conceals its aggregation-prone regions by maintaining a tight fold under crowded conditions. Fusion of NT to the unstructured amyloid-forming A beta(40) peptide, on the other hand, results in the formation of fibrillar aggregates. However, HDX-MS indicates that the NT domain is only partially incorporated into these aggregates in vivo. We conclude that NT is able to control its aggregation to remain functional under the extreme conditions in the spider silk gland.

Authors/Creators:Kaldmae, Margit and Leppert, Axel and Chen, Gefei and Sarr, Medoune and Sahin, Cagla and Nordling, Kerstin and Kronqvist, Nina and Gonzalvo-Ulla, Marta and Fritz, Nicolas and Abelein, Axel and Lain, Sonia and Biverstal, Henrik and Jornvall, Hans and Lane, David P. and Rising, Anna and Johansson, Jan and Landreh, Michael
Title:High intracellular stability of the spidroin N-terminal domain in spite of abundant amyloidogenic segments revealed by in-cell hydrogen/deuterium exchange mass spectrometry
Year of publishing :2020
Volume:287
Page range:2823-2833
Number of Pages:11
Publisher:John Wiley & Sons Ltd, Federation of European Biochemical
ISSN:1742-464X
Language:English
Publication Type:Journal article
Article category:Scientific peer reviewed
Version:Published version
Copyright:Creative Commons: Attribution-Noncommercial 4.0
Full Text Status:Public
Subjects:(A) Swedish standard research categories 2011 > 1 Natural sciences > 106 Biological Sciences (Medical to be 3 and Agricultural to be 4) > Biochemistry and Molecular Biology
Keywords:hydrogen, deuterium exchange mass spectrometry, intracellular protein folding, protein aggregation, spider silk
URN:NBN:urn:nbn:se:slu:epsilon-p-103365
Permanent URL:
http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-p-103365
Additional ID:
Type of IDID
DOI10.1111/febs.15169
Web of Science (WoS)000503637300001
ID Code:17388
Faculty:VH - Faculty of Veterinary Medicine and Animal Science
Department:(VH) > Dept. of Anatomy, Physiology and Biochemistry
Deposited By: SLUpub Connector
Deposited On:02 Sep 2020 09:58
Metadata Last Modified:02 Sep 2020 09:58

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