PURPOSE: To establish a method for studying the proteome of @entity1 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- @entity15 / @entity15 )-based methods for studying the proteome of healthy adult @entity1 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- @entity15 / @entity15 ). In the other method, proteins were first resolved by @entity3208 PAGE ( @entity214 -PAGE) and then in-gel digested with trypsin, and the peptides were analyzed by RPLC/ @entity15 / @entity15 . 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/ @entity15 / @entity15 , and 24 were detected only by @entity214 -PAGE-RPLC- @entity15 / @entity15 . CONCLUSIONS: The data clearly indicated that the @entity214 -PAGE-RPLC- @entity15 / @entity15 method was more suitable than the SCX-RPLC- @entity15 / @entity15 method for analyzing lens fiber proteome. This work greatly expanded the proteome database of @entity1 lens fibers, and the results provided a reference for future studies to detect aging-related and @entity231 -related changes in @entity1 lens fibers proteins.

Comparison of two tandem mass spectrometry-based methods for analyzing the proteome of healthy XXXX lens fibers.

@entity1 :: ('9606', 'Species') :: ['human'] @entity231 :: ('MESH:D002386', 'Disease') :: ['cataract'] @entity3208 :: ('MESH:D012967', 'Chemical') :: ['sodium dodecyl sulfate'] @entity15 :: ('MESH:D009103', 'Disease') :: ['MS'] @entity214 :: ('CHEBI:8984', 'Chemical') :: ['SDS']
@entity1 :: (9606,Species) :: ['human']
