[The analysis of healthy adult male lens fibers proteome using tandem mass spectrometry].
OBJECTIVE
To establish the method for studying the proteome of the lens fibers of healthy adult male in order to provide a tool for further analysis of cataract-related protein using reverse tandem mass spectrometry.


METHODS
Lens fibers proteins were separated and analyzed using sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by RPLC-MS/MS (SDS-PAGE-RPLC/MS/MS). The experiment was repeated three times. The tandem mass spectra of positive results were quality controlled by SEQUEST/AMASS softwares which were based on high abundance ion matching rate, continuity of matched ions and randomnicity score proteins were identified based on 2-hit criteria with reliability over 98% for each peptide.


RESULTS
Total 60 lens fibers proteins were identified at least twice in three experiments. Among these proteins, 22 were identified in previous lens proteome study.


CONCLUSIONS
The results in this study provided preliminary data on the composition of crystallin in the lens fibers of healthy adult male. The results can be used as a reference for future studies to detect the changes of human lens proteins during aging and cataract formation. Combination of SDS-PAGE and RPLC/MS/MS is a quick and simple method, it may be useful for large scale analysis of clinical samples.
Zhang
Chun-wei
CW
Department of Ophthalmology, the First Hospital Affiliated to Harbin Medical University, Harbin 150001, China.
Liu
Ping
P
Wang
Lin-jie
LJ
D000328:Adult; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007908:Lens, Crystalline / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry
D020543:Proteome