Post-translational modification of crystallins in vitreous body from experimental autoimmune uveitis of rats
Post-translational modification of crystallins in vitreous body from experimental autoimmune uveitis of rats
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DOI：
10.1021/pr070133k
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发表时间：
2007-10-01
影响因子：
4.4
通讯作者：
Chung, Hun-Taeg
中科院分区：
文献类型：
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作者：
Bahk, Song-Chul;Jang, Jung-Un;Chung, Hun-Taeg
实验性自身免疫性葡萄膜炎(EAU)是一种广为人知的后葡萄膜炎动物模型，是致盲的主要原因之一。在易感动物(即Lewis大鼠)中，可以通过使用进化上保守的视网膜蛋白(如光受体间视黄醇结合蛋白(IRBP)或该蛋白的表位)进行免疫反应来诱导EAU。首先，我们准备了以下四个试验组，它们随后增加或减少了炎症。(1)正常对照组，(2)IRBP葡萄膜炎组，(3)氯化高铁血红素治疗葡萄膜炎组，(4)二氯原卟啉(SnPP)治疗葡萄膜炎组。其次，采用双向凝胶电泳法(2-DE)、MALDI-TOF/MS和Micro LC/LC-MS/MS分析了Lewis大鼠玻璃体中的渗透蛋白。最后，用Western blotting确定晶状体蛋白的相对含量和(αB晶状体蛋白)的磷酸化位点。在玻璃体中发现了30个斑点，其中27个斑点属于晶体蛋白家族成员。与β-A4和B2-晶体蛋白不同(在未截断的情况下显著增加)，(αA-和B-晶体蛋白仅在EAU玻璃体中被截断。总体而言，在大鼠EAU模型中，我们认为αA和αB晶状体蛋白的翻译后截断、αB晶状体蛋白的磷酸化以及新产生的βA4和βB2晶状体蛋白与葡萄膜炎进展和炎症反应相互关联。
Experimental autoimmune uveitis (EAU) is a well-known animal model of posterior uveitis that is one of the major causes of blindness. EAU could be induced in susceptible animals (i.e., Lewis rat) by immune reactions using evolutionarily conserved retinal proteins, such as interphoto-receptor retinoid binding protein (IRBP), or epitaphs of the protein. First, we prepared the following four test groups that subsequently increased or decreased inflammation. (1) Normal control group, (2) IRBP-induced uveitis group, (3) Hemin-treated uveitis group, and (4) Sn(IV) protoporphyrin IX dichloride (SnPP)treated uveitis group. Second, in the vitreous bodies of Lewis rats, the infiltrated proteins were analyzed using two-dimensional electrophoresis (2-DE), MALDI-TOF/MS, and Micro LC/LC-MS/MS analysis. Finally, Western blotting was applied to confirm the relative amount of crystallins and phosphorylation sites of (alpha B-crystallin. Thirty spots were identified in vitreous bodies, and 27 of these spots were members of the crystallin family. Unlike beta A4- and B2-crystallins (that were significantly increased without truncation), (alpha A- and B-crystallins were only truncated in EAU vitreous body. Taken as a whole, in the rat EAU model, we suggest that post-translational truncations of (alpha A- and alpha B-crystallins, phosphorylation of alpha B-crystallin, and new production of beta A4- and beta B2-crystallins are intercorrelated with uveitis progression and inflammatory responses.