Large-scale identification of human biliary proteins from a cholesterol stone patient using a proteomic approach.
Gallbladder bile, one of the most important body fluids, is composed of water, inorganic ions, conjugated bile salts, phospholipids, cholesterol, bilirubin, mucin and proteins. The separation and identification of bile proteins remain difficult due to the complexity of this matrix. In the present study, human gallbladder bile was obtained from a cholesterol stone patient, and the proteins were isolated and purified by dialysis, precipitation and delipidation procedures. The resulting proteins were divided into several aliquots. One aliquot was subjected to two-dimensional gel electrophoresis (2DE). The protein spots were then in-gel digested and analyzed by liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS). Another aliquot was directly digested and analyzed by a combination of strong cation-exchange (SCX) and reversed-phase (RP) chromatography prior to tandem mass spectrometry (2D-LC/MS/MS). Eventually, 48 and 218 unique proteins were identified from 2DE/MS and 2D-LC/MS/MS, respectively, resulting in a total of 222 unique identified proteins. Of the 218 proteins identified by 2D-LC/MS/MS, 92 were identified based on more than one unique tryptic peptide, and, of the total 222 proteins, 98 were identified based on more than one unique tryptic peptide.
Zhou
Hu
H
Research Centre for Proteome Analysis, Key Lab of Proteomics, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, China.
Chen
Bin
B
Li
Rong-Xia
RX
Sheng
Quan-Hu
QH
Li
Su-Jun
SJ
Zhang
Lei
L
Li
Long
L
Xia
Qi-Chang
QC
Wang
Hong-Yang
HY
Zeng
Rong
R
D000368:Aged; D001646:Bile / Q000737:chemistry* / Q000378:metabolism; D041761:Cholecystolithiasis / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D042882:Gallstones / Q000737:chemistry*; D006801:Humans; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D040901:Proteomics; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
D011506:Proteins