CLONING OF A BRAIN PROTEIN IDENTIFIED BY AUTOANTIBODIES FROM A PATIENT WITH PARA-NEOPLASTIC CEREBELLAR DEGENERATION
CLONING OF A BRAIN PROTEIN IDENTIFIED BY AUTOANTIBODIES FROM A PATIENT WITH PARA-NEOPLASTIC CEREBELLAR DEGENERATION
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DOI：
10.1073/pnas.84.13.4552
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发表时间：
1987-07-01
影响因子：
11.1
通讯作者：
OLD, LJ
中科院分区：
文献类型：
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作者：
DROPCHO, EJ;CHEN, YT;OLD, LJ
在一些副肿瘤性小脑变性患者中已经发现了针对神经元蛋白的自身抗体。为了鉴定这些自身抗体的分子靶标，我们构建了来自人小脑的cDNA表达文库，并用来自副肿瘤性小脑变性患者的IgG筛选该文库。单个克隆pCDR 2产生与患者IgG强烈反应的融合蛋白。分离的pCDR 2克隆用于鉴定6个重叠cDNA克隆。pCDR克隆的测序揭示了由沿整个cDNA序列沿着重复的18个核苷酸（6个氨基酸）的单元组成的独特模式。该序列不同于任何先前描述的真核基因。Southern印迹分析与人类和小鼠基因组中CDR（小脑变性相关）基因的单拷贝表达一致。正常组织的RNA印迹研究显示CDR基因的表达主要限于脑。在源自神经外胚层、肾和肺来源的癌症的细胞系中也注意到CDR信息的表达。
Autoantibodies directed against neuronal proteins have been identified in some patients with paraneoplastic cerebellar degeneration. To identify the molecular targets for these autoantibodies, we constructed a .lambda.gt11 cDNA expression library from human cerebellum and screened the library with IgG from a patient with paraneoplastic cerebellar degeneration. A single clone, pCDR2, produced a fusion protein that reacted strongly with the patient''s IgG. The isolated pCDR2 clone was used to identify six overlapping cDNA clones. Sequencing of the pCDR clones revealed a distinctive pattern consisting of a unit of 18 nucleotides (6 amino acids) repeated in tandem along the entire cDNA sequence. This sequence is unlike any previously described eukaryotic gene. Southern blot analysis was consistent with single-copy representation of the CDR (cerebellum degeneration-related) gene in the human and mouse genome. RNA blotting studies with normal tissues showed expression of the CDR gene to be largely restricted to brain. Expression of the CDR message was also noted in cell lines derived from cancers of neuroectodermal, kidney, and lung origin.