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Simultaneous Quantification of Protein Expression and Modifications by Top-down Targeted Proteomics: A Case of the Sarcomeric Subproteome

Year: 2018
Type: article
Abstract: Determining changes in protein expression and post-translational modifications (PTMs) is crucial for elucidating cellular signal transduction and disease mechanisms. Conventional antibody-based approaches have inherent problems such as the limited availability of high-quality antibodies and batch-to-batch variation. Top-down mass spectrometry (MS)-based proteomics has emerged as the most powerful method for characterization and quantification of protein modifications. Nevertheless, robust methods to simultaneously determine changes in protein expression and PTMs remain lacking. Herein, we have developed a straightforward and robust top-down liquid chromatography (LC)/MS-based targeted proteomics platform for simultaneous quantification of protein expression and PTMs with high throughput and high reproducibility. We employed this method to analyze the sarcomeric subproteome from various muscle types of different species, which successfully revealed skeletal muscle heterogeneity and cardiac developmental changes in sarcomeric protein isoform expression and PTMs. As demonstrated, this targeted top-down proteomics platform offers an excellent 'antibody-independent' alternative for the accurate quantification of sarcomeric protein expression and PTMs concurrently in complex mixtures, which is generally applicable to different species and various tissue types.
DOI: 10.1074/mcp.tir118.001086
Authors: Ziqing Lin, Liming Wei, Wenxuan Cai, Yanlong Zhu, Trisha Tucholski, Stanford D. Mitchell, Wei Guo, Stephen P. Ford, Gary M. Diffee, Ying Ge
Language: English
Subfield: Spectroscopy
Field: Chemistry
Open Access status: hybrid
Estimated APC paid: $3,430
Funders: National Institutes of Health, National Institute of Food and Agriculture, National Heart, Lung, and Blood Institute, NIH Office of the Director, National Institute of General Medical Sciences, Eunice Kennedy Shriver National Institute of Child Health and Human Development
Awards: G1037145829, G1594803720, G3514457801, Cortisol regulation of perinatal adipose tissue and sheep neonatal leptin peak, MASH Explorer, a Comprehensive Software Environment for Top-Down Proteomics, G4286539634, Enabling Top-Down Proteomics through Material Chemistry and Nanotechnology, Deciphering Myofilament Modifications in Ischemic Cardiomyopathy, Training Program In Molecular AND Cellular Pharmacology, Top-Down Proteomics of Myofilaments in Heart Failure, Chemistry-Biology Interface Training Program, Ultra High Resolution Mass Spectrometer for Biomedical Research
OpenAlex ID: W2907850070
FWCI: 1.328
Cites: 72
Cited by: 36
Related to: 10