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J Proteome Res. 2012 Aug 3;11(8):4231-9. doi: 10.1021/pr300358k. Epub 2012 Jul 10.

Human cornea proteome: identification and quantitation of the proteins of the three main layers including epithelium, stroma, and endothelium.

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1
Interdisciplinary Nanoscience Center (iNANO) and Department of Molecular Biology, Aarhus University, and Department of Ophthalmology, Aalborg Hospital, Denmark.

Abstract

Diseases of the cornea are common and refer to conditions like infections, injuries and genetic defects. Morphologically, many corneal diseases affect only certain layers of the cornea and separate analysis of the individual layers is therefore of interest to explore the basic molecular mechanisms involved in corneal health and disease. In this study, the three main layers including, the epithelium, stroma and endothelium of healthy human corneas were isolated. Prior to analysis by LC-MS/MS the proteins from the different layers were either (i) separated by SDS-PAGE followed by in-gel trypsinization, (ii) in-solution digested without prior protein separation or, (iii) in-solution digested followed by cation exchange chromatography. A total of 3250 unique Swiss-Prot annotated proteins were identified in human corneas, 2737 in the epithelium, 1679 in the stroma, and 880 in the endothelial layer. Of these, 1787 proteins have not previously been identified in the human cornea by mass spectrometry. In total, 771 proteins were quantified, 157 based on in-solution digestion and 770 based on SDS-PAGE separation followed by in-gel digestion of excised gel pieces. Protein analysis showed that many of the identified proteins are plasma proteins involved in defense responses.

PMID:
22698189
PMCID:
PMC3411198
DOI:
10.1021/pr300358k
[Indexed for MEDLINE]
Free PMC Article
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