Open AccessThe transient expression of recombinant proteins in plant cell packs facilitates stable isotope labelling for NMR spectroscopy
Author(s) -
Opdensteinen Patrick,
Sperl Laura E.,
Mohamadi Mariam,
KündgenRedding Nicole,
Hagn Franz,
Buyel Johannes Felix
Publication year - 2022
Publication title -
plant biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.525
H-Index - 115
eISSN - 1467-7652
pISSN - 1467-7644
DOI - 10.1111/pbi.13873
Subject(s) - escherichia coli , nuclear magnetic resonance spectroscopy , recombinant dna , labelling , isotope , biochemistry , stable isotope ratio , biology , bacteria , stable isotope labeling by amino acids in cell culture , biophysics , chemistry , proteomics , stereochemistry , physics , quantum mechanics , gene , genetics
Summary Nuclear magnetic resonance (NMR) spectroscopy can be used to determine the structure, dynamics and interactions of proteins. However, protein NMR requires stable isotope labelling for signal detection. The cells used for the production of recombinant proteins must therefore be grown in medium containing isotopically labelled substrates. Stable isotope labelling is well established in Escherichia coli , but bacteria are only suitable for the production of simple proteins without post‐translational modifications. More complex proteins require eukaryotic production hosts, but their growth can be impaired by labelled media, thus reducing product yields and increasing costs. To address this limitation, we used media supplemented with isotope‐labelled substrates to cultivate the tobacco‐derived cell line BY‐2, which was then cast into plant cell packs (PCPs) for the transient expression of a labelled version of the model protein GB1. Mass spectrometry confirmed the feasibility of isotope labelling with 15 N and 2 H using this approach. The resulting NMR spectrum featured a signal dispersion comparable to recombinant GB1 produced in E. coli . PCPs therefore offer a rapid and cost‐efficient alternative for the production of isotope‐labelled proteins for NMR analysis, especially suitable for complex proteins that cannot be produced in microbial systems.