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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >EXPRESSION CLONING OF A HUMAN POLYSIALYLTRANSFERASE THAT FORMS THE POLYSIALYLATED NEURAL CELL ADHESION MOLECULE PRESENT IN EMBRYONIC BRAIN
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EXPRESSION CLONING OF A HUMAN POLYSIALYLTRANSFERASE THAT FORMS THE POLYSIALYLATED NEURAL CELL ADHESION MOLECULE PRESENT IN EMBRYONIC BRAIN

机译：形成人类脑中聚乙烯基化神经细胞粘附分子的人类聚乙烯基转移酶的表达克隆

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Polysialic acid is a developmentally regulated posttranslational modification of the neural cell adhesion molecule (N-CAM). It has been suggested that this large anionic carbohydrate modulates the adhesive property of N-CAM, but the precise function of polysialic acid is not known, Here we describe the isolation and functional expression of a cDNA encoding a human polysialyltransferase. For this expression cloning, COS-1 cells were cotransfected with a human fetal brain cDNA library and a cDNA encoding human N-CAM. Transfected COS-1 cells were stained with a monoclonal antibody specific for polysialic acid and enriched by fluorescence-activated cell sorting, Sibling selection of recovered plasmids resulted in a cDNA clone that directs the expression of polysialic acid on the cell surface, The deduced amino acid sequence indicates that the polysialyltransferase shares a common sequence motif with other sialyltransferases cloned so far. The polysialyltransferase is, however, distinct by having two clusters of basic amino acids, The amount of the polysialyltransferase transcripts correlates well with the formation of polysialic acid in various human tissues, and is abundant in the fetal brain but not in the adult brain. Moreover, HeLa cells stably expressing polysialic acid and N-CAM promoted neurite outgrowth and sprouting. These results indicate that the cloned polysialyltransferase forms polysialylated, embryonic N-CAM, which is critical for plasticity of neural cells. [References: 37]
机译：聚唾液酸是神经细胞粘附分子（N-CAM）的发育调控翻译后修饰。已经有人提出，这种大的阴离子碳水化合物可调节N-CAM的粘附性能，但是聚唾液酸的精确功能尚不清楚。在这里，我们描述了编码人聚唾液酸转移酶的cDNA的分离和功能表达。对于该表达克隆，将COS-1细胞与人胎脑cDNA文库和编码人N-CAM的cDNA共转染。用对聚唾液酸具有特异性的单克隆抗体对转染的COS-1细胞进行染色，并通过荧光激活的细胞分选进行富集。对回收质粒的同胞选择导致了cDNA克隆，该克隆指导聚唾液酸在细胞表面的表达，推导的氨基酸序列表明聚唾液酸转移酶与迄今为止克隆的其他唾液酸转移酶具有共同的序列基序。然而，聚唾液酸转移酶的区别在于具有两个碱性氨基酸簇。聚唾液酸转移酶转录物的量与多种唾液酸在各种人体组织中的形成密切相关，并且在胎儿脑中含量很高，而在成年人脑中含量很高。此外，稳定表达聚唾液酸的HeLa细胞和N-CAM促进神经突生长和发芽。这些结果表明，克隆的聚唾液酸转移酶形成了聚唾液酸化的胚胎N-CAM，这对于神经细胞的可塑性至关重要。 [参考：37]

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