title: Identification of protein N-termini using TMPP or dimethyl labeling and mass spectrometry.
abstract: Determination of a protein's N-terminal sequence can be important for the characterization of protein processing. To increase the confidence of protein N-terminal identification, chemical derivatization of the N-terminal amine group by (N-Succinimidyloxycarbonylmethyl)tris(2,4,6-trimethoxyphenyl)phosphonium bromide (TMPP) or dimethyl labeling followed by mass spectrometric analysis is commonly performed. Using this approach, proteins can be separated by SDS-PAGE, and the protein N-terminus of interest is labeled by TMPP or dimethyl in-gel before tryptic digestion and LC-MS analysis. The N-terminus of a protein can thus be easily identified because the N-terminal tryptic peptides are preferentially labeled. Peptides with N-terminal derivatization produce a better fragmentation pattern during tandem mass spectrometric analysis, which significantly facilitates sequencing of these peptides.
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, Kimmel Center for Biology and Medicine at the Skirball Institute, New York University School of Medicine, New York, NY, 10016, USA.
authors.forename: Jingjing
authors.initials: J
authors.lastname: Deng
authors.forename: Guoan
authors.initials: G
authors.lastname: Zhang
authors.forename: Fang-Ke
authors.initials: FK
authors.lastname: Huang
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Neubert
mesh_terms: D013058:Mass Spectrometry / Q000379:methods; D009943:Organophosphorus Compounds / Q000737:chemistry*; D054730:Protein Interaction Domains and Motifs; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D013194:Staining and Labeling / Q000379:methods
chemical_list: D009943:Organophosphorus Compounds; D011506:Proteins; C405845:tris(trimethoxyphenyl)phosphonium acetyl
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