Nurminskaya Maria V, Talbot Christopher J, Nurminsky Dmitry I, Beazley Kelly E, Linsenmayer Thomas F
Department of Anatomy and Cell Biology, Tufts University, Boston, MA 02111, USA.
Invest Ophthalmol Vis Sci. 2009 Aug;50(8):3655-61. doi: 10.1167/iovs.08-3170. Epub 2009 Feb 28.
Ferritin is an iron storage protein that is generally cytoplasmic. However, in embryonic avian corneal epithelial (CE) cells, the authors previously observed that the ferritin was predominantly nuclear. They also obtained evidence that this ferritin protects DNA from oxidative damage by UV light and hydrogen peroxide and that the nuclear localization involves a tissue-specific nuclear transporter, termed ferritoid. In the present investigation, the authors have determined additional properties of the nuclear ferritoid-ferritin complexes.
For biochemical characterization, a combination of molecular sieve chromatography, immunoblotting, and nuclear-cytoplasmic fractionation was used; DNA binding was analyzed by electrophoretic mobility shift assay.
The CE nuclear ferritin complex has characteristics that differentiate it from a "typical" cytoplasmic ferritin, including the presence of ferritin and ferritoid subunits; a molecular weight of approximately 260 kDa, which is approximately half that of cytoplasmic ferritin; its iron content, which is below our limits of detection; and its ability to bind to DNA.
Within CE cell nuclei, ferritin and ferritoid are coassembled into stable complex(es) present in embryonic and adult corneas. Thus, ferritoid not only serves transiently as a nuclear transporter for ferritin, it remains as a component of a unique ferritoid-ferritin nuclear complex.
铁蛋白是一种通常存在于细胞质中的铁储存蛋白。然而,作者先前在胚胎期鸡角膜上皮(CE)细胞中观察到,铁蛋白主要存在于细胞核中。他们还获得了证据,表明这种铁蛋白可保护DNA免受紫外线和过氧化氢的氧化损伤,且核定位涉及一种组织特异性核转运蛋白,称为类铁蛋白。在本研究中,作者确定了核类铁蛋白-铁蛋白复合物的其他特性。
为进行生化特性分析,采用了分子筛色谱、免疫印迹和核质分级分离相结合的方法;通过电泳迁移率变动分析来分析DNA结合情况。
CE细胞核铁蛋白复合物具有使其有别于“典型”细胞质铁蛋白的特性,包括存在铁蛋白和类铁蛋白亚基;分子量约为260 kDa,约为细胞质铁蛋白的一半;其铁含量低于我们的检测限;以及其与DNA结合的能力。
在CE细胞核内,铁蛋白和类铁蛋白共同组装成存在于胚胎和成年角膜中的稳定复合物。因此,类铁蛋白不仅暂时作为铁蛋白的核转运蛋白,它还作为独特的类铁蛋白-铁蛋白核复合物的一个组成部分保留下来。