Proteome profiling of wild type and lumican-deficient mouse corneas
Proteome profiling of wild type and lumican-deficient mouse corneas
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DOI：
10.1016/j.jprot.2011.04.032
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发表时间：
2011-09-06
影响因子：
3.3
通讯作者：
Chakravarti, Shukti
中科院分区：
文献类型：
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作者：
Shao, HanJuan;Chaerkady, Raghothama;Chakravarti, Shukti
为了阐明一种主要的角膜蛋白多糖LUMICAN的缺失如何影响角膜的动态平衡，我们使用质谱仪得出了LUMICAN缺陷和杂合子小鼠角膜的蛋白质组图谱，并将其与野生型角膜蛋白质组进行了比较。2108个蛋白质在小鼠的角膜中被定量。所选蛋白质和转录本分别用Western印迹和定量RT-PCR进行检测。我们观察到在发光剂缺乏的小鼠体内基质细胞外基质(ECM)蛋白的组成发生了重大变化。Lumcan缺乏症改变了基质和角膜上皮中的细胞蛋白。Lum(+/-)和Lum(-)小鼠角膜的ECM改变包括形成I型胶原、VI型胶原、纤维调素、Perlecan、层粘连蛋白82、IV型胶原、Nidogen/entactin和锚定的VII型胶原的纤维增加，而Lum(-)角膜基质中的蛋白多糖、二聚糖和角化多糖减少。细胞蛋白质的变化包括酒精脱氢酶、超氧化物歧化酶的增加以及上皮细胞角蛋白8和14的减少。我们还检测到了对角膜来说是新的蛋白质。这些蛋白质组将提供对基质变薄和透明度丧失的发光蛋白缺乏的角膜表型的洞察，并更好地了解角膜和眼营养不良的致病变化。(C)2011爱思唯尔B.V.保留所有权利。
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 82, 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. (C) 2011 Elsevier B.V. All rights reserved.