Human cornea proteome: identification and quantitation of the proteins of the three main layers including epithelium, stroma, and endothelium.
Human cornea proteome: identification and quantitation of the proteins of the three main layers including epithelium, stroma, and endothelium.
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DOI：
10.1021/pr300358k
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发表时间：
2012-08-03
影响因子：
4.4
通讯作者：
Enghild JJ
中科院分区：
文献类型：
--
作者：
Dyrlund TF;Poulsen ET;Scavenius C;Nikolajsen CL;Thøgersen IB;Vorum H;Enghild JJ
角膜疾病是常见的，是指感染、损伤和遗传缺陷等疾病。在形态学上，许多角膜疾病仅影响角膜的某些层，因此对各个层的单独分析对于探索涉及角膜健康和疾病的基本分子机制是有意义的。本研究分离了健康人角膜的上皮、基质和内皮三个主要层。在通过LC-MS/MS分析之前，将来自不同层的蛋白质（i）通过SDS-PAGE分离，随后进行凝胶内胰蛋白酶消化，（ii）在没有预先蛋白质分离的情况下进行溶液内消化，或（iii）在溶液内消化，随后进行阳离子交换色谱。在人角膜中共鉴定了3250种独特的Swiss-Prot注释蛋白，在上皮中鉴定了2737种，在基质中鉴定了1679种，在内皮层中鉴定了880种。其中，1787种蛋白质此前尚未通过质谱法在人类角膜中鉴定出来。总共对771种蛋白质进行了定量，157种基于溶液中消化，770种基于SDS-PAGE分离，然后对切下的凝胶片进行凝胶内消化。蛋白质分析表明，许多鉴定的蛋白质是参与防御反应的血浆蛋白。
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.
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影响因子：
4.2
作者：
Feener, Edward P.
通讯作者：
Feener, Edward P.
影响因子：
4.4
作者：
Blais, DR;Vascotto, SG;Altosaar, I
通讯作者：
Altosaar, I
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作者：
Ayala, A;Warejecka, DJ;Yue, BYJT
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Yue, BYJT
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作者：
Chowdhury, Uttio Roy;Madden, Benjamin J.;Fautsch, Michael P.
通讯作者：
Fautsch, Michael P.
影响因子：
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作者：
Jha, Purushottam;Bora, Puran S.;Bora, Nalini S.
通讯作者：
Bora, Nalini S.