Sedimentation Yields Long-Term Stable Protein Samples as Shown by Solid-State NMR.
nucleotides
proteins
sedimentation
solid-state NMR
stability
Journal
Frontiers in molecular biosciences
ISSN: 2296-889X
Titre abrégé: Front Mol Biosci
Pays: Switzerland
ID NLM: 101653173
Informations de publication
Date de publication:
2020
2020
Historique:
received:
18
12
2019
accepted:
30
01
2020
entrez:
11
3
2020
pubmed:
11
3
2020
medline:
11
3
2020
Statut:
epublish
Résumé
Today, the sedimentation of proteins into a magic-angle spinning (MAS) rotor gives access to fast and reliable sample preparation for solid-state Nuclear Magnetic Resonance (NMR), and this has allowed for the investigation of a variety of non-crystalline protein samples. High protein concentrations on the order of 400 mg/mL can be achieved, meaning that around 50-60% of the NMR rotor content is protein; the rest is a buffer solution, which includes counter ions to compensate for the charge of the protein. We have demonstrated herein the long-term stability of four sedimented proteins and complexes thereof with nucleotides, comprising a bacterial DnaB helicase, an ABC transporter, an archaeal primase, and an RNA polymerase subunit. Solid-state NMR spectra recorded directly after sample filling and up to 5 years later indicated no spectral differences and no loss in signal intensity, allowing us to conclude that protein sediments in the rotor can be stable over many years. We have illustrated, using an example of an ABC transporter, that not only the structure is maintained, but that the protein is still functional after long-term storage in the sedimented state.
Identifiants
pubmed: 32154263
doi: 10.3389/fmolb.2020.00017
pmc: PMC7047159
doi:
Types de publication
Journal Article
Langues
eng
Pagination
17Informations de copyright
Copyright © 2020 Wiegand, Lacabanne, Torosyan, Boudet, Cadalbert, Allain, Meier and Böckmann.
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