MOLECULAR-CLONING OF A HUMAN PRION PROTEIN CDNA
MOLECULAR-CLONING OF A HUMAN PRION PROTEIN CDNA
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DOI：
10.1089/dna.1986.5.315
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发表时间：
1986-08-01
期刊：
影响因子：
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通讯作者：
DEARMOND, SJ
中科院分区：
文献类型：
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作者：
KRETZSCHMAR, HA;STOWRING, LE;DEARMOND, SJ
人的克鲁兹-雅克病(CJD)和动物的瘙痒病是由普恩病毒引起的退行性、传播性神经系统疾病。已知的唯一的大分子内的蛋白是蛋白(PrP)。仓鼠Prion蛋白(PrP27-30)的编码基因已被克隆和测序(Oesch等人，1985)。利用该仓鼠PrP基因，我们筛选了人视网膜cDNA文库，并对杂交插入片段最长的cDNA克隆进行了测序。该插入片段含有一个编码人蛋白的长开放阅读框架(ORF)。Northern转移分析表明，相关的Poly(A)+RNA测量.apprx。2.5kb在多种人类神经外胚层细胞系中表达。人PrP与仓鼠PrP在253个氨基酸中有27个不同，在759个ORF核苷酸中有98个不同。仓鼠和人PrP氨基酸序列的保守率接近90%，反映了两种蛋白质相似的结构特征和共同的抗原性(Bockman等人，1985)。人PrP序列包含一个由22个氨基酸组成的氨基端信号肽，两个足以跨膜的疏水片段，以及两个可能的N-糖基化位点。仓鼠和人类Prion蛋白之间的保守性表明，它们可能在细胞新陈代谢中发挥重要作用，并可能解释瘙痒病和CJD之间的相似之处。
Cruetzfeldt-Jakob disease (CJD) of humans and scrapie of animals are degenerative, transmissible neurologic diseases caused by prions. The only known macromolecules within prions are prion proteins (PrP). The cDNA encoding the hamster prion protein (PrP 27-30) has been cloned and sequenced (Oesch et al., 1985). Using that hamster PrP cDNA, we screened a human retina cDNA library and sequenced the cDNA clone with the longest hybridizing insert. This insert was found to contain a long open reading frame (ORF) encoding the human prion protein. Northern transfer analysis showed that a related poly(A)+RNA measuring .apprx. 2.5 kb is expressed in a variety of human neuroectodermal cell lines. Human PrP differed from hamster PrP at 27 of 253 amino acids and at 98 of 759 ORF nucleotides. Conservation of PrP amino acid sequence between hamster and human is nearly 90%, reflecting similar structural features and shared antigenicity of the two proteins (Bockman et al., 1985). The human PrP sequence contained a presumptive amino-terminal signal peptide of 22 amino acids, two hydrophobic segments of sufficient length to span membranes, and two possible sites for N-glycosylation. The conservation between the hamster and human prion protein suggests that they may have an important role in cellular metabolism and may explain the similarities between scrapie and CJD.