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 st...

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Bibliographic Details
Published in:Rapid communications in mass spectrometry 2005-01, Vol.19 (23), p.3569-3578
Main Authors: Zhou, Hu, Chen, Bin, Li, Rong-Xia, Sheng, Quan-Hu, Li, Su-Jun, Zhang, Lei, Li, Long, Xia, Qi-Chang, Wang, Hong-Yang, Zeng, Rong
Format: Article
Language:English
Subjects:
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Bile - chemistry
Bile - metabolism
Cholecystolithiasis - metabolism
Chromatography, High Pressure Liquid
Electrophoresis, Gel, Two-Dimensional
Female
Gallstones - chemistry
Humans
Proteins - chemistry
Proteins - metabolism
Proteomics
Spectrometry, Mass, Electrospray Ionization - methods
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Summary: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. Copyright © 2005 John Wiley & Sons, Ltd.
ISSN:0951-4198
1097-0231
DOI:10.1002/rcm.2207