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Quantification of Protein Isoforms in Mesenchymal Stem Cells by Reductive Dimethylation of Lysines in Intact Proteins by Yi-Min She; Michael Rosu-Myles; Lisa Walrond; Terry D. Cyr is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Quantification of Protein Isoforms in Mesenchymal Stem Cells by Reductive Dimethylation of Lysines in Intact Proteins about?

Mass spectrometry (MS)-based quantification of highly homologous proteins in complex samples has proven difficult due to subtle sequence variations and the wide dynamic range of protein isoforms present. Herein, we report the use of reductive dimethylation on intact proteins to quantitatively compare protein isoform expression in the nucleus and cytoplasm of mesenchymal stem cells (MSC) and normal stroma. By coupling fixed-charge MS/MS scanning, high-resolution UPLC FT-MS data-dependent acquisition and MASCOT-based data mining, hydrogen/deuterium-labeled dimethyl-lysine peptides were simultaneously captured allowing the accurate comparison of 123 protein isoforms in parallel LC MS/MS runs. Thirty-four isoforms were identified that had expression levels specific to MSC. Where possible, proteomic analyses were verified by Western blotting and were demonstrated to be divergent from the level of gene transcription detected for certain proteins. Our analysis provides a protein isoform signature specific to MSC and demonstrates the suitability of dimethyl-lysine labeling on intact proteins for quantifying highly homologous proteins on a proteome-wide scale.

Who reads Quantification of Protein Isoforms in Mesenchymal Stem Cells by Reductive Dimethylation of Lysines in Intact Proteins?

Author

Yi-Min She; Michael Rosu-Myles; Lisa Walrond; Terry D. Cyr

Publisher

John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Ltd.; Wiley; Research Square (ISSN 1615-9853)

Published

2012

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)