Expression cloning of a CMP-NeuAc:NeuAcα2-3Galβ1-4Glcβ1-1′Cer α2,8-sialyltransferase (GD3 synthase) from human melanoma cells

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作者：

Kiyomitsu Nara，Yumiko Watanabe，Kazuo Maruyama，Kohji Kasahara，Yutaka Sanai

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摘要：

Using an expression cloning approach, we have isolated a cDNA encoding GD3 synthase (CMp-NeuAc: NeuAcα2-3Galβ1-4Glcβ 1-1'Cer α 2,8-sialytransferase, EC 2.4.99.8), which is a key regulatory enzyme determining the prominence of the ganglioside biosynthesis pathway. The cloned cDNA encodes a 341-amino acid protein containing a single transmembrane domain at its N-terminal region, suggesting that the protein has a type II transmembrane topology. The sequence of α2,8-sialytransferase showed a high level of similarity with other sialytransferases at two conserved regions typical in the sialytransferase family. transfected cells containing the cloned cDNA expressed GD3 ganglioside on the cell surface, which was detectable with specific anti-GD3 antibody by immunofluorescence and immunostaining after separation of isolated glycolipids on this-layer chromatography. The cDNA hybridized to a single mRNA species of 2.4 kb in melanoma cells. This sialytransferase is distinctive in catalyzing the formation of the α2-8 linkage of sialic acids.

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关键词：

Biochemistry Glycosyltransferase GD3 Ganglioside Tumor Antigen Melanoma

DOI：

10.1073/pnas.91.17.7952

被引量：

202

年份：

1994