ArticlePlant methods2006
Metabolic labeling of plant cell cultures with K(15)NO3 as a tool for quantitative analysis of proteins and metabolites.
Article in Plant methods, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed.
- Stable Isotope-Assisted Plant Metabolomics: Combination of Global and Tracer-Based Labeling for Enhanced Untargeted Profiling and Compound Annotation.Frontiers in plant science · 2019Article
- Advanced Proteomic Approaches to Elucidate Somatic Embryogenesis.Frontiers in plant science · 2018Review
- Identification of Cargo for Adaptor Protein (AP) Complexes 3 and 4 by Sucrose Gradient Profiling.Molecular & cellular proteomics : MCP · 2016Article
- Protein Phosphorylation and Redox Modification in Stomatal Guard Cells.Frontiers in physiology · 2016Review
- Changes in the Phosphoproteome and Metabolome Link Early Signaling Events to Rearrangement of Photosynthesis and Central Metabolism in Salinity and Oxidative Stress Response in Arabidopsis.Plant physiology · 2015Article
- The role of proteomics in progressing insights into plant secondary metabolism.Frontiers in plant science · 2015Review
- Development of a GC/Quadrupole-Orbitrap mass spectrometer, part II: new approaches for discovery metabolomics.Analytical chemistry · 2014Article
- Metabolic labeling and membrane fractionation for comparative proteomic analysis of Arabidopsis thaliana suspension cell cultures.Journal of visualized experiments : JoVE · 2013Article
- Quantification of histone modifications using ¹⁵N metabolic labeling.Methods (San Diego, Calif.) · 2013Review
- Quantitative phosphoproteomics after auxin-stimulated lateral root induction identifies an SNX1 protein phosphorylation site required for growth.Molecular & cellular proteomics : MCP · 2013Article
- Plant SILAC: stable-isotope labelling with amino acids of arabidopsis seedlings for quantitative proteomics.PloS one · 2013Article
- iMS2Flux--a high-throughput processing tool for stable isotope labeled mass spectrometric data used for metabolic flux analysis.BMC bioinformatics · 2012Article
- Precision, proteome coverage, and dynamic range of Arabidopsis proteome profiling using (15)N metabolic labeling and label-free approaches.Molecular & cellular proteomics : MCP · 2012Article
- Production of isotopically labeled standards from a uniformly labeled precursor for quantitative volatile metabolomic studies.Analytical chemistry · 2012Article
- Determining degradation and synthesis rates of arabidopsis proteins using the kinetics of progressive 15N labeling of two-dimensional gel-separated protein spots.Molecular & cellular proteomics : MCP · 2012Article
- Proteome dynamics and early salt stress response of the photosynthetic organism Chlamydomonas reinhardtii.BMC genomics · 2012Article
- Gel-based and gel-free quantitative proteomics approaches at a glance.International journal of plant genomics · 2012Article
- Quantitation, networking, and function of protein phosphorylation in plant cell.Frontiers in plant science · 2012Article
- Quantitative shotgun proteomics using a uniform ¹⁵N-labeled standard to monitor proteome dynamics in time course experiments reveals new insights into the heat stress response of Chlamydomonas reinhardtii.Molecular & cellular proteomics : MCP · 2011Article
- Quantitative proteomics by metabolic labeling of model organisms.Molecular & cellular proteomics : MCP · 2010Article
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Strategies for robust quantitative comparison between different biological samples are of high importance in experiments that address biological questions beyond the establishment of protein lists. Here, we propose the use of 15N-KNO3 as the only nitrogen source in Arabidopsis cell cultures in order to achieve a metabolically fully labeled cell population. Proteins from such metabolically labeled culture are distinguishable from unlabeled protein populations by a characteristic mass shift that depends on the amino acid composition of the tryptic peptide analyzed. In addition, the metabolically labeled cell extracts are also suitable for comparative quantitative analysis of nitrogen-containing cellular metabolic complement. Protein extracts from unlabeled and from standardized 15N-labeled cells were combined into one sample for joined analytical processing. This has the advantage of (i) reduced experimental variability and (ii) immediate relative quantitation at the level of single extracted peptide and metabolite spectra. Together ease and accuracy of relative quantitation for profiling experiments is substantially improved. The metabolic labeling strategy has been validated by mixtures of protein extracts and metabolite extracts from the same cell cultures in known ratios of labeled to unlabeled extracts (1:1, 1:4, and 4:1). We conclude that saturating metabolic 15N-labeling provides a robust and affordable integrative strategy to answer questions in quantitative proteomics and nitrogen focused metabolomics.
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Registered trials
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