Proteome profiling of wild type and lumican-deficient mouse corneas

• Published in: Journal of proteomics Vol. 74; no. 10; pp. 1895 - 1905
• Main Authors: Shao, HanJuan, Chaerkady, Raghothama, Chen, Shoujun, Pinto, Sneha M., Sharma, Rakesh, Delanghe, Bernard, Birk, David E., Pandey, Akhilesh, Chakravarti, Shukti
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier B.V 06-09-2011; Elsevier
• Subjects: alcohol dehydrogenase; Animals; Biological and medical sciences; Chondroitin Sulfate Proteoglycans - deficiency; Chondroitin Sulfate Proteoglycans - genetics; Collagen; Cornea; Cornea - metabolism; Diverse techniques; epithelium; Epithelium, Corneal - metabolism; Epithelium, Corneal - pathology; extracellular matrix; Extracellular Matrix Proteins - metabolism; Fundamental and applied biological sciences. Psychology; Gene Expression Profiling; heterozygosity; homeostasis; iTRAQ; Keratan Sulfate - deficiency; Keratan Sulfate - genetics; laminin; Lumican; Mass spectrometry; Mice; Molecular and cellular biology; phenotype; proteoglycans; proteome; Proteome - metabolism; Proteomics; reverse transcriptase polymerase chain reaction; Spectrometry, Mass, Electrospray Ionization; superoxide dismutase; Western blotting; iTRAQ; Cornea; Lumican; Mass spectrometry; Collagen; Proteomics; Vertebrata; Mammalia; Mouse; Rodentia; Proteome
• ISSN: 1874-3919; 1876-7737
• DOI: 10.1016/j.jprot.2011.04.032

To elucidate how the deficiency of a major corneal proteoglycan, lumican, affects corneal homeostasis, we used mass spectrometry to derive the proteome profile of the lumican-deficient and the heterozygous mouse corneas and compared these to the wild type corneal proteome. 2108 proteins were quantified in the mouse cornea. Selected proteins and transcripts were investigated by Western blot and quantitative RT-PCR, respectively. We observed major changes in the composition of the stromal extracellular matrix (ECM) proteins in the lumican-deficient mice. Lumican deficiency altered cellular proteins in the stroma and the corneal epithelium. The ECM changes included increases in fibril forming collagen type I, Collagen type VI, fibromodulin, perlecan, laminin β2, collagen type IV, nidogen/entactin and anchoring collagen type VII in the Lum+/− and the Lum−/− mouse corneas, while the stromal proteoglycans decorin, biglycan and keratocan were decreased in the Lum−/− corneas. Cellular protein changes included increases in alcohol dehydrogenase, superoxide dismutase and decreases in epithelial cytokeratins 8 and 14. We also detected proteins that are novel to the cornea. The proteomes will provide an insight into the lumican-deficient corneal phenotype of stromal thinning and loss of transparency and a better understanding of pathogenic changes in corneal and ocular dystrophies.
Deep coverage of mouse corneal proteome using quantitative proteomics provides insight into functional classes of proteins in the mouse cornea and the main effects of lumican deficiency on corneal homeostasis. [Display omitted]
► Lumican is a major corneal proteoglycan that affects corneal homeostasis. ► An iTRAQ-based quantitative proteomic tool was used to study the Lum–/– mouse cornea. ► Major changes epithelial and stromal proteins were observed.