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 | ||||||
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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 | ||||||
Series Name/Journal: | FEBS Journal | ||||||
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: |
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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: | 15 Jan 2021 19:24 |
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