中文摘要：

背景视神经蛋白（OPTN）基因是青光眼和肌萎缩性脊髓侧索硬化的致病基因，在眼部视网膜组织中的表达最多。我们先前的研究分离出与OPTN相互作用的蛋白质，确定其中之一为视网膜色素变性的致病基因——碱性亮氨酸拉链（bZIP）转录因子，即神经视网膜亮氨酸（NRL）拉链，并证实OPTN和NRL两蛋白间存在相互作用。目的确定与NRL相互作用所必需的OPTN蛋白结合区域。方法构建一系列带FLAG标签的OPTN部分片段缺失质粒，分别与带血凝素标记的NRL（HA—NRL）共转染HeLaS3细胞。用抗HA和抗FLAG标签抗体分别对细胞质和细胞核提取物进行免疫共沉淀和Westernblot检测，分析两蛋白质问的相互结合方式，以确定NRL与OPTN蛋白的结合区域。结果在OPTN的第一、第二和第三缺失区域质粒转染细胞的细胞核组分中均检测出血凝素标记的NRL（HA-NRL）免疫共沉淀条带，但在第四缺失区域质粒中不产生此HA—NRL条带。从所有缺失质粒及全长OPTN质粒转染细胞的细胞质组分中均未观察到NRL条带。结论本研究进一步证实OPTN和NRL在HeLaS3细胞核内的相互作用，并明确其蛋白结合区域。在共表达NRL和OPTN的HeLaS3细胞中，Flag-OPTN可与HA-NRL相互结合，且OPTN的尾端区域（423-577）是结合NRL所必需的。

英文摘要：

Background Gene encoding optineurin （OPTN） is a causative gene for glaucoma and amyotrophic lateral sclerosis,with a more expression in retina. Our previous study isolated OPTN-interacting proteins and identified that the gene encode the basic leucine zipper （bZIP） transcription factor neural retina leucine （NRL） zipper,a causative gene for retinitis pigmentosa, and further study demonstrated the interaction between OPTN and NRL proteins in nuclei of cultured HeLaS3 cells. Objective This study was to determine the protein binding site of OPTN necessary for NRL binding. Methods A deletion series of OPTN-expression plasmids were constructed and co-expressed with hemagglutinin （HA） -tagged NRL in HeLaS3 cells, respectively. The cytoplasmic and nuclear fractions were used to perform co-immunoprecipitate （ColP） and Western blot with anti-tag antibodies. Results In the nuclear fractions of cells transfected with the dell st, del2nd or del3rd plasmid, a band of coimmunoprecipitated HA-labelled NRL （HA-NRL） was detected. However,the del4th plasmid did not produce a band. The NRL band was not found in cytoplasmic fractions from transfected cells with any of the deletion plasmids or with the whole-length OPTN plasmid. Conclusions The protein binding site of OPTN necessary for NRL binding is determined. This result demonstrates the binding of Flag-OPTN and HA-NRL in HeLaS3 cells. A series of partial-deletion OPTN plasmids demonstrated that the tail region （423-577 amino acids） of OPTN was necessary for binding with NRL.