ArticleAnalytical chemistry2011
Quantitative label-free phosphoproteomics strategy for multifaceted experimental designs.
Article in Analytical chemistry, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed, 48 citations in OpenAlex.
- Protein modification by deamidation indicates variations in joint extracellular matrix turnover.The Journal of biological chemistry · 2012Trial
- Regulation of organic anion transporting polypeptide 1B1 transport function by concurrent phosphorylation and lysine-acetylation: A novel posttranslational regulation mechanism.Molecular pharmacology · 2025Article
- Quantitative Phosphoproteomic Comparison of Lens Proteins in Highly Myopic Cataract and Age-Related Cataract.BioMed research international · 2021Article
- Label-free quantitative proteomic analysis identifies CTNNB1 as a direct target of FOXP3 in gastric cancer cells.Oncology letters · 2018Article
- Acylation of Superoxide Dismutase 1 (SOD1) at K122 Governs SOD1-Mediated Inhibition of Mitochondrial Respiration.Molecular and cellular biology · 2017Article
- Analyses of PDE-regulated phosphoproteomes reveal unique and specific cAMP-signaling modules in T cells.Proceedings of the National Academy of Sciences of the United States of America · 2017Article
- The analysis of heterotaxy patients reveals new loss-of-function variants of GRK5.Scientific reports · 2016Article
- The G Protein-Coupled Estrogen Receptor Agonist G-1 Inhibits Nuclear Estrogen Receptor Activity and Stimulates Novel Phosphoproteomic Signatures.Toxicological sciences : an official journal of the Society of Toxicology · 2016Article
- S-Nitrosylation of Sarcomeric Proteins Depresses Myofilament Ca2+)Sensitivity in Intact Cardiomyocytes.Antioxidants & redox signaling · 2015Article
- Label-free quantitative phosphoproteomics with novel pairwise abundance normalization reveals synergistic RAS and CIP2A signaling.Scientific reports · 2015Article
- Quantifying reversible oxidation of protein thiols in photosynthetic organisms.Journal of the American Society for Mass Spectrometry · 2015Article
- Single-step enrichment by Ti4+-IMAC and label-free quantitation enables in-depth monitoring of phosphorylation dynamics with high reproducibility and temporal resolution.Molecular & cellular proteomics : MCP · 2014Article
- Label-free quantitation of phosphopeptide changes in erythrocyte membranes: towards molecular mechanisms underlying deformability alterations in stored red blood cells.Haematologica · 2014Article
- Evaluation of quantitative performance of sequential immobilized metal affinity chromatographic enrichment for phosphopeptides.Analytical biochemistry · 2014Article
- Phosphoproteomic profiling of human myocardial tissues distinguishes ischemic from non-ischemic end stage heart failure.PloS one · 2014Article
- Longitudinal study of differential protein expression in an Alzheimer's mouse model lacking inducible nitric oxide synthase.Journal of proteome research · 2013Article
- Identification and functional analysis of phosphorylation residues of the Arabidopsis BOTRYTIS-INDUCED KINASE1.Protein & cell · 2013Article
- Solid-phase capture for the detection and relative quantification of S-nitrosoproteins by mass spectrometry.Methods (San Diego, Calif.) · 2013Review
- Proteomic analysis of human bronchoalveolar lavage fluid after subsgemental exposure.Journal of proteome research · 2013Article
- Biochemical and biophysical changes underlie the mechanisms of basement membrane disruptions in a mouse model of dystroglycanopathy.Matrix biology : journal of the International Society for Matrix Biology · 2013Article
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Authors and funding
5 authors at 3 institutions in 1 country.
Funding
Abstract
Protein phosphorylation is a critical regulator of signaling in nearly all eukaryotic cellular pathways and dysregulated phosphorylation has been implicated in an array of diseases. The majority of MS-based quantitative phosphorylation studies are currently performed from transformed cell lines because of the ability to generate large amounts of starting material with incorporated isotopically labeled amino acids during cell culture. Here we describe a general label-free quantitative phosphoproteomic strategy capable of directly analyzing relatively small amounts of virtually any biological matrix, including human tissue and biological fluids. The strategy utilizes a TiO(2) enrichment protocol in which the selectivity and recovery of phosphopeptides were optimized by assessing a twenty-point condition matrix of binding modifier concentrations and peptide-to-resin capacity ratios. The quantitative reproducibility of the TiO(2) enrichment was determined to be 16% RSD through replicate enrichments of a wild-type Danio rerio (zebrafish) lysate. Measured phosphopeptide fold-changes from alpha-casein spiked into wild-type zebrafish lysate backgrounds were within 5% of the theoretical value. Application to a morpholino induced knock-down of G protein-coupled receptor kinase 5 (GRK5) in zebrafish embryos resulted in the quantitation of 719 phosphorylated peptides corresponding to 449 phosphorylated proteins from 200 μg of zebrafish embryo lysates.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.