TY - JOUR
T1 - Proteome profiling of wild type and lumican-deficient mouse corneas
AU - Shao, Han Juan
AU - Chaerkady, Raghothama
AU - Chen, Shoujun
AU - Pinto, Sneha M.
AU - Sharma, Rakesh
AU - Delanghe, Bernard
AU - Birk, David E.
AU - Pandey, Akhilesh
AU - Chakravarti, Shukti
PY - 2011/9/6
Y1 - 2011/9/6
N2 - 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 β 2, 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.
AB - 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 β 2, 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.
KW - Collagen
KW - Cornea
KW - ITRAQ
KW - Lumican
KW - Mass spectrometry
KW - Proteomics
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U2 - 10.1016/j.jprot.2011.04.032
DO - 10.1016/j.jprot.2011.04.032
M3 - Article
C2 - 21616181
AN - SCOPUS:80052036854
SN - 1874-3919
VL - 74
SP - 1895
EP - 1905
JO - Journal of Proteomics
JF - Journal of Proteomics
IS - 10
ER -