Chapter title |
Identification of Protein N-Termini Using TMPP or Dimethyl Labeling and Mass Spectrometry
|
---|---|
Chapter number | 19 |
Book title |
Proteomic Profiling
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2550-6_19 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2549-0, 978-1-4939-2550-6
|
Authors |
Jingjing Deng, Guoan Zhang, Fang-Ke Huang, Thomas A. Neubert |
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. |
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