机构：[1]Department of Ophthalmology, the First Affiliated Hospital, Harbin Medical University, Harbin, China[2]Beijing Tong Ren Eye Center, Beijing Tong Ren Hospital, Capital Medical University, Beijing, China首都医科大学附属北京同仁医院首都医科大学附属同仁医院[3]Proteomics Research Center, National Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences/Peking Union Medical College, Beijing, China
出处：
ISSN：
摘要：
Purpose: To establish a method for studying the proteome of human lens fibers and to provide the proteome database of lens fibers from healthy male adults. Methods: We compared two liquid chromatography tandem mass spectrometry (LC-MS/MS)-based methods for studying the proteome of healthy adult human lens fibers. Total proteins were extracted from pooled lens fibers of 12 healthy male adult donors. In one method, the total proteins were digested with trypsin, and the derived peptides were analyzed by strong cation exchange (SCX) coupled with reverse phase liquid chromatography tandem mass spectrometry (RPLC-MS/MS). In the other method, proteins were first resolved by sodium dodecyl sulfate PAGE (SDS-PAGE) and then in-gel digested with trypsin, and the peptides were analyzed by RPLC/MS/MS. The tandem mass spectra of positive results were quality controlled by advanced mass spectrum scanner (AMASS) software. The peptide false positive rate was estimated using the reverse database searching method. Results: A total of 68 proteins from lens fibers were identified using these two methods based on at least two different peptide matches with reliability of over 97% for each peptide. Among these proteins, 43 were detected by both methods, one was detected only by SCX-RPLC/MS/MS, and 24 were detected only by SDS-PAGE-RPLC-MS/MS. Conclusions: The data clearly indicated that the SDS-PAGE-RPLC-MS/MS method was more suitable than the SCX-RPLC-MS/MS method for analyzing lens fiber proteome. This work greatly expanded the proteome database of human lens fibers, and the results provided a reference for future studies to detect aging-related and cataract-related changes in human lens fibers proteins.
基金：
This work was supported in part by grants from the Science
and Technology Foundation of the Department of Education,
Heilong Jiang Province (No:11521162); Foundation for Key
Problems in Science and Technology, Heilong Jiang province
(No:GB03C602-1); and Foundation for the Innovation of
Graduate Students, Heilong Jiang Province
(No:SCX2005038).
第一作者机构：[1]Department of Ophthalmology, the First Affiliated Hospital, Harbin Medical University, Harbin, China
通讯作者：
通讯机构：[1]Department of Ophthalmology, the First Affiliated Hospital, Harbin Medical University, Harbin, China[*1]23 You Zheng Street, Nan Gang District, Department of Ophthalmology, The First Affiliated Hospital, Harbin Medical University, Harbin, People’s Republic of China
推荐引用方式(GB/T 7714)：
Zhang Chunwei,Liu Ping,Wang Ningli,et al.Comparison of two tandem mass spectrometry-based methods for analyzing the proteome of healthy human lens fibers[J].MOLECULAR VISION.2007,13(209-10):1873-1877.
APA：
Zhang, Chunwei,Liu, Ping,Wang, Ningli,Li, Yaojun&Wang, Linjie.(2007).Comparison of two tandem mass spectrometry-based methods for analyzing the proteome of healthy human lens fibers.MOLECULAR VISION,13,(209-10)
MLA：
Zhang, Chunwei,et al."Comparison of two tandem mass spectrometry-based methods for analyzing the proteome of healthy human lens fibers".MOLECULAR VISION 13..209-10(2007):1873-1877
