PXD064210

PXD064210 is an original dataset announced via ProteomeXchange.

Title	Optimization of Bio-Orthogonal Non-Canonical Amino acid Tagging (BONCAT) for effective low-disruption labelling of Arabidopsis proteins in vivo
Description	Plants, as sessile organisms, require robust responses in protein synthesis to adapt to variable environmental conditions. Measurement of newly synthesized proteins has been successfully facilitated with Bio-Orthogonal Non-Canonical Amino acid Tagging (BONCAT) across a multitude of organisms. Here, we use non-canonical amino acids (NCAAs) L-azidohomoalanine (AHA) or L-homopropargylglycine (HPG) incorporation in place of methionine residues into the actively translating Arabidopsis proteome, allowing for that subset of proteins to be enriched for mass spectrometry quantification. Although this technique has seen occasional use in plants, optimization of the protocol to maximize functionality while minimizing organismal stress has not yet been established. Here, we provide evidence for successful implementation through the liquid immersion of seedlings in AHA or HPG-containing media that functions with significantly lower concentrations than the literature standard. Our approach splits acute exposure and incorporation phase of labelling to mitigate potential negative impacts of prolonged NCAA exposure without compromising effective enrichment capacity, and demonstrate that this results in an unperturbed growth phenotype for AHA-treated seedlings. Finally, we demonstrate the capacity of this modified approach to enrich newly synthesized proteins from the whole proteome under standard stress conditions. These improvements allow for a broader use of BONCAT technologies in molecular plant research, affording a deeper understanding of the newly synthesized proteome without negatively impacting plant health.
HostingRepository	PRIDE
AnnounceDate	2026-06-08
AnnouncementXML	Submission_2026-06-07_16:30:05.479.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Richard Uhrig
SpeciesList	scientific name: Arabidopsis thaliana (Mouse-ear cress); NCBI TaxID: NEWT:3702;
ModificationList	biotinylated residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2025-05-22 15:05:48	ID requested
⏵ 1	2026-06-07 16:30:06	announced

Hassan N, Braun S, Talasila M, Kennedy C, Yaremko L, Fahlman RP, Uhrig RG, Optimizing bio-orthogonal non-canonical amino acid tagging (BONCAT) for low-disruption labeling of Arabidopsis proteins in vivo. Plant Physiol, 201(1):(2026) [pubmed]

10.1093/plphys/kiaf573;

submitter keyword: Mass Spectrometry,BONCAT, Click Chemistry, Quantitative Proteomics

R. Glen Uhrig
contact affiliation	Department of Biological Sciences, University of Alberta
contact email	ruhrig@ualberta.ca
lab head
Richard Uhrig
contact affiliation	University of Alberta
contact email	ruhrig@ualberta.ca
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064210

PRIDE project URI

• PRIDE
  1. PXD064210
     1. Label: PRIDE project
     2. Name: Optimization of Bio-Orthogonal Non-Canonical Amino acid Tagging (BONCAT) for effective low-disruption labelling of Arabidopsis proteins in vivo