[The analysis of healthy adult male lens fibers proteome using tandem mass spectrometry]

Abstract

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.

Conclusion: 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.