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HeeJung Ju, Jeewon Mok, Chang Rae Rho, Ji Young Kwon, Choun-Ki Joo; Expression Profiling of Keratoconic Cornea using MALDI-TOF mass spectrometry. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):5489.
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© ARVO (1962-2015); The Authors (2016-present)
To identify proteins differentially expressed between the cornea of keratoconus(KC) patient and control subject using two dimensional electrophoresis (2-DE) and mass spectrometry-based techniques
Proteins from the whole cornea were isolated from keratoconus and normal cornea tissues, and separated by two-dimensional gel electrophoresis (2-DE). Comparison of protein patterns was performed using PDQuest Software and protein differences were identified 100 differential protein spots by mass spectrometry. All of the 100 protein spots were analyzed the peptide mass finger printing (PMF) with the 4700 MALDI-TOF/TOF mass spectrometer to deduce their possible functions. Finally, results were validated by western-blot
Of the 100 protein spots, we identified 51 spots that demonstrated a significant differences between keratoconus and reference normal cornea with a ProtScore (>or=2). Particularly, the intensity of 37 spots were markedly higher in keratoconus than normal cornea (>2 folds), and 14 spots were detected only in keratoconic cornea. Among them, transforming growth factor-beta-induced protein ig-h3 precursor, Keratin 14, and alpha-crystallin B chain were more expressed in keratoconus pateints and aldehyde dehydrogenase 1, keratin 3, alpha-enolase and peroxiredoxin-6 were more expressed in control
In this study, we have presented the finding of proteins differentially expressed in the keratoconus patients. The identified proteins corresponded to molecules belonging to ECM related cytoskeleton, antioxidant protein and transcription/transformation. Our findings could contribute to the knowledge for of pathogenesis of keratoconus
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