Stable isotope labeling method targeting terminal tyrosine for relative peptide quantitation using mass spectrometry

被引:6
作者
Choi, Yong Seok [1 ]
Pennington, Christopher L. [1 ]
Wood, Troy D. [1 ]
机构
[1] SUNY Buffalo, Dept Chem, Buffalo, NY 14260 USA
关键词
Stable isotope labeling; Peptide quantitation; Neuropeptide; Tyrosine; Tyrosinase; Cysteine; LC-MS; CEREBROSPINAL-FLUID; PROTEIN EXPRESSION; QUANTIFICATION; BIOMARKERS; PROTEOMICS; MIXTURES;
D O I
10.1016/j.ab.2010.02.027
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Many neuropeptides lack suitable amino acid residues for modification by existing selective isotope labeling methods and use in relative quantitation by mass spectrometry. To address this issue, a new stable isotope labeling method that targets tyrosine residues by coupling with light cysteine (do) or heavy cysteine (d(2)) in the presence of tyrosinase was developed. Optimal derivatization conditions fort 1 mu M leucine-enkephalin were achieved when 10 mM cysteine and 200 U/ml tyrosinase at pH 6.8 to 7.2 were used for a 60-min incubation period at room temperature. Under these conditions, leucine-enkephalin present at concentrations as low as 125 nM was successfully labeled. When comparisons between the lightly labeled (do) and heavily labeled (d(2)) forms were made, a discrepancy between the actual concentration ratio and the raw peak intensity ratio was observed; this is due to the overlap of an isotopic peak of the do with the monoisotopic peak of d(2). Fortunately, this discrepancy can be corrected by one of two simple computational approaches described. The quantitative labeling of this method to neuropeptides with the terminal tyrosine was confirmed and provides an alternative when other selective isotope-coded affinity tagging methods are not suitable. (C) 2010 Elsevier Inc. All rights reserved.
引用
收藏
页码:15 / 21
页数:7
相关论文
共 37 条
[1]   Quantitative mass spectrometry in proteomics: a critical review [J].
Bantscheff, Marcus ;
Schirle, Markus ;
Sweetman, Gavain ;
Rick, Jens ;
Kuster, Bernhard .
ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2007, 389 (04) :1017-1031
[2]   Proteomic evaluation of core biopsy specimens from breast lesions [J].
Bisca, A ;
D'Ambrosio, C ;
Scaloni, A ;
Puglisi, F ;
Aprile, G ;
Piga, A ;
Zuiani, C ;
Bazzocchi, M ;
Di Loreto, C ;
Paron, I ;
Tell, G ;
Damante, G .
CANCER LETTERS, 2004, 204 (01) :79-86
[3]   Practical implications of some recent studies in electrospray ionization fundamentals [J].
Cech, NB ;
Enke, CG .
MASS SPECTROMETRY REVIEWS, 2001, 20 (06) :362-387
[4]   8-Plex quantitation of changes in cerebrospinal fluid protein expression in subjects undergoing intravenous immunoglobulin treatment for Alzheimer's disease [J].
Choe, Leila ;
D'Ascenzo, Mark ;
Relkin, Norman R. ;
Pappin, Darryl ;
Ross, Philip ;
Williamson, Brian ;
Guertin, Steven ;
Pribil, Patrick ;
Lee, Kelvin H. .
PROTEOMICS, 2007, 7 (20) :3651-3660
[5]   Silanization of inner surfaces of nanoelectrospray ionization emitters for reduced analyte adsorption [J].
Choi, Yong Seok ;
Wood, Troy D. .
RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 2008, 22 (08) :1265-1274
[6]   Quantification of Gluten Exorphin A5 in cerebrospinal fluid by liquid chromatography-mass spectrometry [J].
Fanciulli, G ;
Azara, E ;
Wood, TD ;
Dettorl, A ;
Delitala, G ;
Marchetti, M .
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, 2006, 833 (02) :204-209
[7]   Liquid chromatography-mass spectrometry assay for quantification of Gluten Exorphin B5 in cerebrospinal fluid [J].
Fanciulli, Giuseppe ;
Azara, Emanuela ;
Wood, Troy D. ;
Delitala, Giuseppe ;
Marchetti, Mauro .
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, 2007, 852 (1-2) :485-490
[8]   Cerebrospinal fluid proteomic biomarkers for Alzheimer's disease [J].
Finehout, Erin J. ;
Franck, Zsofia ;
Choe, Leila H. ;
Relkin, Norman ;
Lee, Kelvin H. .
ANNALS OF NEUROLOGY, 2007, 61 (02) :120-129
[9]   Absolute quantification of proteins and phosphoproteins from cell lysates by tandem MS [J].
Gerber, SA ;
Rush, J ;
Stemman, O ;
Kirschner, MW ;
Gygi, SP .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2003, 100 (12) :6940-6945
[10]   Stable isotopic labeling in proteomics [J].
Gevaert, Kris ;
Impens, Francis ;
Ghesquiere, Bart ;
Van Damme, Petra ;
Lambrechts, Anja ;
Vandekerckhove, Joel .
PROTEOMICS, 2008, 8 (23-24) :4873-4885