Mass-balanced H-1/H-2 isotope dipeptide tag for simultaneous protein quantitation and identification
SCIE
SCOPUS
- Title
- Mass-balanced H-1/H-2 isotope dipeptide tag for simultaneous protein quantitation and identification
- Authors
- SEO, JONGCHEOL; Suh, MS; Thangadurai, TD; Kim, J; Rhee, YH; Yoon, HJ; Shin, SK
- Date Issued
- 2008-08-15
- Publisher
- AMER CHEMICAL SOC
- Abstract
- Mass-balanced H-1/H-2 isotope dipeptide tags (MBITs) are presented for simultaneous protein quantitation and identification. MBIT is derived from N-acetyl-Ala-Ala dipeptide and conjugated to primary amines of target peptides. H-1/H-2 isotopes are encoded in the methyl groups of N-acetylated dipeptide: one tag deuterated on the N-acetyl group and another on the C-terminal alanine. MBIT-linked peptides comigrate in reversed-phase liquid chromatography without significant H-1/H-2 isotope effects and provide 2-plex quantitation signals at 114 and 117 Th as well as peptide sequence information upon MS/MS analysis with MALDI TOF/TOF. MBIT shows good quantitation linearity in a concentration range of 20-250 fmol. The performance of MBIT on protein quantitation and identification is further tested with yeast heat-shock protein (Hsp82p) obtained from three different physiological states. MBIT using nanogram-scale samples produces the relative abundance ratios comparable to those obtained from optical imaging of microgram-scale samples visualized with SYPRO Ruby stain. The MBIT strategy is a simple and low-cost alternative for 2-plex quantitation of proteins and offers possibilities of tuning the 2-plex signal mass window by replacing the N-terminal alanine with other amino acid residues.
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/29414
- DOI
- 10.1021/AC801007Y
- ISSN
- 0003-2700
- Article Type
- Article
- Citation
- ANALYTICAL CHEMISTRY, vol. 80, no. 16, page. 6145 - 6153, 2008-08-15
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