ArticleMethods in molecular biology (Clifton, N.J.)2016
Proteomic Analysis of Protein Turnover by Metabolic Whole Rodent Pulse-Chase Isotopic Labeling and Shotgun Mass Spectrometry Analysis.
Article in Methods in molecular biology (Clifton, N.J.), 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed.
- Simultaneous profiling of native-state proteomes and transcriptomes of neural cell types using proximity labeling.Nature communications · 2026Article
- Reduction of RAD23A extends lifespan and mitigates pathology in a mouse model of TDP-43 proteinopathy.Nature communications · 2026Article
- Exosome Proteomics of SOD1Annals of clinical and translational neurology · 2026Article
- Utilizing Omic Data to Understand Integrative Physiology.Physiology (Bethesda, Md.) · 2025Review
- Local mitochondrial replication in the periphery of neurons requires the eEF1A1 protein and thetranslation of nuclear-encoded proteins.iScience · 2024Article
- Circadian gene expression in mouse renal proximal tubule.American journal of physiology. Renal physiology · 2023Article
- An atlas of protein turnover rates in mouse tissues.Nature communications · 2021Article
- Using stable isotope labeling to advance our understanding of Alzheimer's disease etiology and pathology.Journal of neurochemistry · 2021Review
- Long-lived mitochondrial cristae proteins in mouse heart and brain.The Journal of cell biology · 2021Article
- Pulse-Chase Proteomics of the App Knockin Mouse Models of Alzheimer's Disease Reveals that Synaptic Dysfunction Originates in Presynaptic Terminals.Cell systems · 2021Article
- Isotopic Nitrogen-15 Labeling of Mice Identified Long-lived Proteins of the Renal Basement Membranes.Scientific reports · 2020Article
- Exploring ribosome composition and newly synthesized proteins through proteomics and potential biomedical applications.Expert review of proteomics · 2017Review
- Metaproteomics as a Complementary Approach to Gut Microbiota in Health and Disease.Frontiers in chemistry · 2017Review
Corrections and comments
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Authors and funding
3 authors.
Funding
Abstract
The analysis of protein half-life and degradation dynamics has proven critically important to our understanding of a broad and diverse set of biological conditions ranging from cancer to neurodegeneration. Historically these protein turnover measures have been performed in cells by monitoring protein levels after "pulse" labeling of newly synthesized proteins and subsequent chase periods. Comparing the level of labeled protein remaining as a function of time to the initial level reveals the protein's half-life. In this method we provide a detailed description of the workflow required for the determination of protein turnover rates on a whole proteome scale in vivo. Our approach starts with the metabolic labeling of whole rodents by restricting all the nitrogen in their diet to exclusively nitrogen-15 in the form of spirulina algae. After near complete organismal labeling with nitrogen-15, the rodents are then switched to a normal nitrogen-14 rich diet for time periods of days to years. Tissues are harvested, the extracts are fractionated, and the proteins are digested to peptides. Peptides are separated by multidimensional liquid chromatography and analyzed by high resolution orbitrap mass spectrometry (MS). The nitrogen-15 containing proteins are then identified and measured by the bioinformatic proteome analysis tools Sequest, DTASelect2, and Census. In this way, our metabolic pulse-chase approach reveals in vivo protein decay rates proteome-wide.
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