ArticleJournal of proteome research2012
GeLC-MRM quantitation of mutant KRAS oncoprotein in complex biological samples.
Article in Journal of proteome research, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 37 citations in OpenAlex.
- Polysomal Profiling Coupled to Allele-Specific Proteomics Reveals an EIF4H TranSNP Allele Possessing Higher mRNA Translation Potential.Molecular & cellular proteomics : MCP · 2026Article
- Detection and Quantitation of Endogenous Membrane-Bound RAS Proteins and KRAS Mutants in Cancer Cell Lines Using 1D-SDS-PAGE LC-MSMethods in molecular biology (Clifton, N.J.) · 2024Article
- GLOBAL AND TARGETED PROFILING OF GTP-BINDING PROTEINS IN BIOLOGICAL SAMPLES BY MASS SPECTROMETRY.Mass spectrometry reviews · 2021Review
- Hemoglobin oxidation generates globin-derived peptides in atherosclerotic lesions and intraventricular hemorrhage of the brain, provoking endothelial dysfunction.Laboratory investigation; a journal of technical methods and pathology · 2020Article
- Relative stabilities of wild-type and mutant glial fibrillary acidic protein in patients with Alexander disease.The Journal of biological chemistry · 2019Article
- Roles of Small GTPases in Acquired Tamoxifen Resistance in MCF-7 Cells Revealed by Targeted, Quantitative Proteomic Analysis.Analytical chemistry · 2018Article
- A Targeted Quantitative Proteomic Approach Assesses the Reprogramming of Small GTPases during Melanoma Metastasis.Cancer research · 2018Article
- Quantification of mutant SPOP proteins in prostate cancer using mass spectrometry-based targeted proteomics.Journal of translational medicine · 2017Article
- A Timely Shift from Shotgun to Targeted Proteomics and How It Can Be Groundbreaking for Cancer Research.Frontiers in oncology · 2017Review
- Oncogenic KRAS and BRAF Drive Metabolic Reprogramming in Colorectal Cancer.Molecular & cellular proteomics : MCP · 2016Article
- Review
- The clinical impact of recent advances in LC-MS for cancer biomarker discovery and verification.Expert review of proteomics · 2016Review
- Evaluating kinase ATP uptake and tyrosine phosphorylation using multiplexed quantification of chemically labeled and post-translationally modified peptides.Methods (San Diego, Calif.) · 2015Article
- Absolute Quantification of Endogenous Ras Isoform Abundance.PloS one · 2015Article
- Onco-proteogenomics: cancer proteomics joins forces with genomics.Nature methods · 2014Article
- Identification and Quantitation of Coding Variants and Isoforms of Pulmonary Surfactant Protein A.Journal of proteome research · 2014Article
- Demonstrating the feasibility of large-scale development of standardized assays to quantify human proteins.Nature methods · 2014Article
- Proteomic approaches in biomarker discovery: new perspectives in cancer diagnostics.TheScientificWorldJournal · 2014Review
- Connecting genomic alterations to cancer biology with proteomics: the NCI Clinical Proteomic Tumor Analysis Consortium.Cancer discovery · 2013Article
- Proteomic analysis of exosomes from mutant KRAS colon cancer cells identifies intercellular transfer of mutant KRAS.Molecular & cellular proteomics : MCP · 2013Article
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
Tumor-derived mutant KRAS (v-Ki-ras-2 Kirsten rat sarcoma viral oncogene) oncoprotein is a critical driver of cancer phenotypes and a potential biomarker for many epithelial cancers. Targeted mass spectrometry analysis by multiple reaction monitoring (MRM) enables selective detection and quantitation of wild-type and mutant KRAS proteins in complex biological samples. A recently described immunoprecipitation approach (Proc. Nat. Acad. Sci.2011, 108, 2444-2449) can be used to enrich KRAS for MRM analysis, but requires large protein inputs (2-4 mg). Here, we describe sodium dodecyl sulfate-polyacrylamide gel electrophoresis-based enrichment of KRAS in a low molecular weight (20-25 kDa) protein fraction prior to MRM analysis (GeLC-MRM). This approach reduces background proteome complexity, thus, allowing mutant KRAS to be reliably quantified in low protein inputs (5-50 μg). GeLC-MRM detected KRAS mutant variants (G12D, G13D, G12V, G12S) in a panel of cancer cell lines. GeLC-MRM analysis of wild-type and mutant was linear with respect to protein input and showed low variability across process replicates (CV = 14%). Concomitant analysis of a peptide from the highly similar HRAS and NRAS proteins enabled correction of KRAS-targeted measurements for contributions from these other proteins. KRAS peptides were also quantified in fluid from benign pancreatic cysts and pancreatic cancers at concentrations from 0.08 to 1.1 fmol/μg protein. GeLC-MRM provides a robust, sensitive approach to quantitation of mutant proteins in complex biological samples.
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