Frank M B, McCubbin V R, Heldermon C
Arthritis and Immunology Program, Oklahoma Medical Research Foundation, Oklahoma City.
Biochem J. 1995 Jan 15;305 ( Pt 2)(Pt 2):359-62. doi: 10.1042/bj3050359.
The 52 kDa Ro/SSA protein is an intracellular autoantigen that is frequently recognized by antibodies in sera of patients with systemic lupus erythematosus or Sjogren's syndrome. While the function of this molecule is not known, zinc finger and leucine zipper motifs have been identified in its predicted amino acid sequence which suggest that it may interact with nucleic acids. To test this hypothesis, the human gene which encodes this protein was cloned in a baculovirus and expressed in Spodoptera frugipoda cells. Extracts from these infected insect cells were used as a source of protein for this study. The protein is similar in size and antigenicity to that expressed in human cells. This protein binds to DNA at physiological temperature and is eluted with high concentrations of sodium chloride. Striking similarities were found between the sequence in, and adjacent to, the nucleic acid-binding motifs of 52 kDa Ro/SSA and a growing family of zinc finger proteins which have been shown to bind to DNA or regulate gene expression. The findings presented here place this protein structurally and functionally in this family and demonstrate a biochemical assay which can be used to study its function.
52 kDa Ro/SSA蛋白是一种细胞内自身抗原,系统性红斑狼疮或干燥综合征患者血清中的抗体常常能识别该抗原。虽然此分子的功能尚不清楚,但在其预测的氨基酸序列中已鉴定出锌指和亮氨酸拉链基序,这表明它可能与核酸相互作用。为验证这一假说,编码该蛋白的人类基因被克隆到杆状病毒中,并在草地贪夜蛾细胞中表达。来自这些感染昆虫细胞的提取物用作本研究的蛋白质来源。该蛋白在大小和抗原性上与在人类细胞中表达的蛋白相似。这种蛋白在生理温度下与DNA结合,并能用高浓度氯化钠洗脱。在52 kDa Ro/SSA的核酸结合基序及其相邻序列与一个不断扩大的锌指蛋白家族之间发现了惊人的相似性,这些锌指蛋白已被证明能与DNA结合或调节基因表达。本文给出的研究结果从结构和功能上把这种蛋白归到这个家族中,并展示了一种可用于研究其功能的生化检测方法。