Localization of the human 64kD autoantigen D1 to myofibrils in a subset of extraocular muscle fibers
Localization of the human 64kD autoantigen D1 to myofibrils in a subset of extraocular muscle fibers
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DOI：
10.1076/ceyr.19.4.313.5304
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发表时间：
1999-10-01
影响因子：
2
通讯作者：
Fowler, VM
中科院分区：
文献类型：
--
作者：
Conley, CA;Fowler, VM
目的。探讨64kD人类自身抗原d1的组织特异性表达模式，该蛋白是一种可能参与甲状腺相关眼病的促甲状腺激素相关蛋白。重组64kD人自身抗原D_1在细菌表达系统中表达，免疫兔。对正常大鼠、甲亢大鼠和兔组织进行免疫印迹，并对大鼠组织冰冻切片进行免疫荧光定位。抗64kD人类自身抗原D1抗体可特异性识别眼外肌、胸骨甲状腺肌和平滑肌蛋白印迹中与70kD相似的多肽。免疫荧光染色显示64kD人类自身抗原D1定位于大鼠眼外肌和胸骨甲状腺肌慢纤维中的肌原纤维。甲亢兔眼外肌中该蛋白的表达水平无明显变化。64kD的人类自身抗原D1作为肌原纤维的一部分在眼外肌和胸骨甲状腺肌的慢纤维中表达，在甲状腺功能亢进的情况下不上调。
Purpose. To evaluate the tissue-specific expression pattern of the 64kD human autoantigen D1, a tropomodulin-related protein that may be involved in thyroid-associated ophthalmopathy.Methods. Recombinant 64kD human autoantigen D1 was generated in a bacterial expression system and used to immunize rabbits. Specific antibodies were affinity-purified and used for Western blots on normal and hyperthyroid rat and rabbit tissue, and immunofluorescence localization on cryosections of rat tissue.Results. Anti-64kD human autoantigen D1 antibodies recognize specifically a similar to 70kD polypeptide in western blots of extraocular muscle, sternothyroid muscle, and smooth muscle. Immunofluorescence staining demonstrates that the 64kD human autoantigen D1 localizes to myofibrils in slow fibers from rat extraocular and sternothyroid muscle. The level of this protein is not altered in extraocular muscles from hyperthyroid rabbits.Conclusions. The 64kD human autoantigen D1 is expressed in slow fibers of extraocular and sternothyroid muscles as a component of myofibrils, and is not upregulated in conditions of hyperthyroidism.