title: N-terminal isotope tagging with propionic anhydride: proteomic analysis of myogenic differentiation of C2C12 cells.
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 C2C12 myoblasts upon their myogensis to myotubules.
authors.affiliation: Department of Biochemistry, College of Science, Protein Network Research Center, Yonsei University, 134 Sinchon-dong, Seoul 120-749, Republic of Korea.
authors.forename: Hyung Wook
authors.initials: HW
authors.lastname: Nam
authors.forename: Richard
authors.initials: R
authors.lastname: Simpson
authors.forename: Yu Sam
authors.initials: YS
authors.lastname: Kim
mesh_terms: D000595:Amino Acid Sequence; D000812:Anhydrides / Q000737:chemistry*; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D003903:Deuterium; D007553:Isotope Labeling / Q000379:methods*; D051379:Mice; D008969:Molecular Sequence Data; D032446:Myoblasts / Q000166:cytology*; D010446:Peptide Fragments / Q000737:chemistry; D011422:Propionates / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*; D012710:Serum Albumin, Bovine; D021241:Spectrometry, Mass, Electrospray Ionization; D014357:Trypsin; D014746:Vimentin / Q000378:metabolism
chemical_list: D000812:Anhydrides; D010446:Peptide Fragments; D011422:Propionates; D014746:Vimentin; D012710:Serum Albumin, Bovine; D003903:Deuterium; C096126:propionic anhydride; D014357:Trypsin