Large-scale identification of proteins in human salivary proteome by liquid chromatography/mass spectrometry and two-dimensional gel electrophoresis-mass spectrometry

被引:293
作者
Hu, S
Xie, YM
Ramachandran, P
Loo, RRO
Li, Y
Loo, JA [1 ]
Wong, DT
机构
[1] Univ Calif Los Angeles, Dept Chem & Biochem, Los Angeles, CA 90095 USA
[2] Univ Calif Los Angeles, Sch Dent, Los Angeles, CA 90024 USA
[3] Univ Calif Los Angeles, Dent Res Inst, Los Angeles, CA 90024 USA
[4] Univ Calif Los Angeles, David Geffen Sch Med, Dept Biol Chem, Los Angeles, CA 90095 USA
[5] Univ Calif Los Angeles, Inst Mol Biol, Los Angeles, CA 90095 USA
[6] Univ Calif Los Angeles, Jonsson Comprehens Canc Ctr, Los Angeles, CA 90024 USA
[7] Univ Calif Los Angeles, Mass Spectrometry & Proteom Ctr, Los Angeles, CA 90095 USA
关键词
two-dimensional gel electrophoresis; Liquid chromatography mass spectrometry; matrix-assisted laser desorption; ionization - mass spectrometry; salivary proteome; shotgun proteomics;
D O I
10.1002/pmic.200401037
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Human saliva contains a large number of proteins and peptides (salivary proteome) that help maintain homeostasis in the oral cavity. Global analysis of human salivary proteome is important for understanding oral health and disease pathogenesis. in this study, large-scale identification of salivary proteins was demonstrated by using shotgun proteomics and two-dimensinal gel electrophoresis-mass spectrometry (2-DE-MS). For the shotgun approach, whole saliva proteins were prefractionated according to molecular weight. The smallest fraction, presumably containing salivary peptides, was directly separated by capillary liquid chromatography (LC). However, the large protein fractions were digested into peptides for subsequent LC separation. Separated peptides were analyzed by on-line electrospray tandem mass spectrometry (MS/MS) using a quadrupole-time of flight mass spectrometer, and the obtained spectra were automatically processed to search human protein sequence database for protein identification. Additionally, 2-DE was used to map out the proteins in whole saliva. Protein spots 105 in number were excised and in-gel digested; and the resulting peptide fragments were measured by matrix-assisted laser desorption/ionization-mass spectrometry and sequenced by LC-MS/MS for protein identification. In total, we cataloged 309 proteins from human whole saliva by using these two proteomic approaches.
引用
收藏
页码:1714 / 1728
页数:15
相关论文
共 37 条
[1]   Toward a human blood serum proteome - Analysis by multidimensional separation coupled with mass spectrometry [J].
Adkins, JN ;
Varnum, SM ;
Auberry, KJ ;
Moore, RJ ;
Angell, NH ;
Smith, RD ;
Springer, DL ;
Pounds, JG .
MOLECULAR & CELLULAR PROTEOMICS, 2002, 1 (12) :947-955
[2]   Mass spectrometry-based proteomics [J].
Aebersold, R ;
Mann, M .
NATURE, 2003, 422 (6928) :198-207
[3]   Cysteine protease inhibitory activity and levels of salivary cystatins in whole saliva of periodontally diseased patients [J].
Baron, AC ;
Gansky, SA ;
Ryder, MI ;
Featherstone, JDB .
JOURNAL OF PERIODONTAL RESEARCH, 1999, 34 (08) :437-444
[4]   Peptide mapping of proteins in human body fluids using electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry [J].
Bergquist, J ;
Palmblad, M ;
Wetterhall, M ;
Håkansson, P ;
Markides, KE .
MASS SPECTROMETRY REVIEWS, 2002, 21 (01) :2-15
[5]   Lectin probe analysis of the glycosylation of human parotid salivary glycoproteins [J].
Carpenter, GH ;
Proctor, GB ;
Pankhurst, CL ;
Zhang, XS ;
Shori, DK .
BIOCHEMICAL SOCIETY TRANSACTIONS, 1995, 23 (04) :S538-S538
[6]  
Carpenter GH, 1999, ELECTROPHORESIS, V20, P2124, DOI 10.1002/(SICI)1522-2683(19990701)20:10<2124::AID-ELPS2124>3.0.CO
[7]  
2-M
[8]   Determination of the human salivary peptides histatins 1, 3, 5 and statherin by high-performance liquid chromatography and by diode-array detection [J].
Castagnola, M ;
Congiu, D ;
Denotti, G ;
Di Nunzio, A ;
Fadda, MB ;
Melis, S ;
Messana, I ;
Misiti, F ;
Murtas, R ;
Olianas, A ;
Piras, V ;
Pittau, A ;
Puddu, G .
JOURNAL OF CHROMATOGRAPHY B, 2001, 751 (01) :153-160
[9]   Analysis of the saliva from patients with oral cancer by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry [J].
Chen, YC ;
Li, TY ;
Tsai, MF .
RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 2002, 16 (05) :364-369
[10]   Mapping of proteins in human saliva using two-dimensional gel electrophoresis and peptide mass fingerprinting [J].
Ghafouri, B ;
Tagesson, C ;
Lindahl, M .
PROTEOMICS, 2003, 3 (06) :1003-1015