N-terminal isotope tagging with propionic anhydride: Proteomic analysis of myogenic differentiation of C2C12 cells

Author(s): Nam, HW; Simpson, R; Kim, YS;
Details: Publication Year 2005-11-05,Volume 826,Issue #1-2,Page 91-107
Journal Title: JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Publication Type: Journal Article
Abstract: N-Terminal isotope tagging (NIT) is an important proteomic tool for quantifying proteins in complex mixtures. Here we describe a modified version of the isotope-coded propionylation procedure of Zhang et al. [Zhang et al., Rapid Commun. Mass Spectom. 16 (2002) 2325], which uses 'light' D0 and 'heavy' D10-propionic anhydride. The method has been extensively modified to improve both the kinetics and overall yield of propionylation. Using albumin as a model protein, the overall variation in quantification yields, calculated using several tryptic peptides, was within +/- 10% (S.D. +/- 0.2) error. The efficacy of the method is demonstrated by the quantitative differences obtained for vimentin in cell lysates of C2CI2 myoblasts upon their myogensis to myotubules. (c) 2005 Elsevier B.V. All rights reserved.
Publisher: ELSEVIER SCIENCE BV
Keywords: TANDEM MASS-SPECTROMETRY; CODED AFFINITY TAGS; QUANTITATIVE PROTEOMICS; LIQUID-CHROMATOGRAPHY; PROTEIN IDENTIFICATION; SHOTGUN PROTEOMICS; AMINO-ACIDS; QUANTIFICATION; TOOLS; ELECTROPHORESIS
Publisher's Version: https://doi.org/10.1016/j.jchromb.2005.07.039
Terms of Use/Rights Notice: Refer to copyright notice on published article.

Creation Date: 2005-11-05 12:00:00