PXD075774-1

PXD075774 is an original dataset announced via ProteomeXchange.

Title	Cupriavidus necator, an alternative source for isotopic enrichment of proteins expressed in insect cells for NMR
Description	Isotopic enrichment of pharmacologically relevant protein targets is crucial for structural studies by nucleIsotopic enrichment of pharmacologically relevant protein targets is crucial for structural studies by nuclear magnetic resonance (NMR) and plays a key role in advancing structure-guided drug discovery. Many clinically important drug targets require expression in eukaryotic systems-such as mammalian, yeast, or insect cells-rather than prokaryotic hosts. This requirement limits the feasibility of high-throughput isotopic labeling and poses challenges for obtaining uniformly isotope-labeled proteins suitable for NMR analysis. While several enrichment strategies have been developed, no broadly applicable enrichment platform has emerged for eukaryotic expression systems. In this study, we introduce Cupriavidus necator as an alternative biological source for 15N and 13C isotopic enrichment to support protein production in eukaryotic systems. To evaluate this approach, we selected the kinase domain of EPHA2, a receptor tyrosine kinase implicated in colorectal cancer progression and an important target for therapeutic inhibitor development. Isotopic incorporation was quantified using liquid chromatography-mass spectrometry (LC-MS), revealing enrichment levels of 79% for 15N and 69% for 13C. These results demonstrate that Cupriavidus necator can serve as a robust and flexible platform for generating isotopically enriched biomolecules compatible with eukaryotic protein expression, thereby enabling NMR investigations of disease-relevant protein targets.
HostingRepository	PRIDE
AnnounceDate	2026-03-26
AnnouncementXML	Submission_2026-03-26_10:04:10.099.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Julian Langer
SpeciesList	scientific name: Nosema spodopterae; NCBI TaxID: NEWT:231490;
ModificationList	acetylated residue; iodoacetamide derivatized residue
Instrument	impact II

Revision	Datetime	Status	ChangeLog Entry
0	2026-03-17 18:09:37	ID requested
⏵ 1	2026-03-26 10:04:10	announced

Dataset with its publication pending

submitter keyword: Cupriavidus necator, isotope labeling, NMR

Julian Langer
contact affiliation	MPI of Biophysics and MPI of Brain Research
contact email	julian.langer@biophys.mpg.de
lab head
Julian Langer
contact affiliation	MPIs for Biophysics and Brain Research
contact email	julian.langer@biophys.mpg.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD075774
     1. Label: PRIDE project
     2. Name: Cupriavidus necator, an alternative source for isotopic enrichment of proteins expressed in insect cells for NMR