Expression cloning of a human alpha l,4-Nacetylglucosaminyltransfe that forms GlcNAc aphal l to 4 Gal beta to R, a glycan specificanlly expressed in the gastric gland mucous cell-type mucin

JUN NAKAYAMA; JIUNN-CHERN YEH; ANUP K.MISKRA

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Expression cloning of a human alpha l,4-Nacetylglucosaminyltransfe that forms GlcNAc aphal l to 4 Gal beta to R, a glycan specificanlly expressed in the gastric gland mucous cell-type mucin

机译：人α1，4-N乙酰基氨基葡萄糖氨基转移蛋白的表达克隆，其形成GlcNAc aphal 1至4 Gal beta至R，一种在胃腺黏液细胞型黏蛋白中特异性表达的聚糖

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Among mucus-secreting cells, the gastric gland mucous cells, Brunnei''s glands, accessory glands of pancreati-cohiliary tract, and pancreatic ducts exhibiting gastric metaplasia are unique in that they express class lll mucin identified by paradoxical Con A staining coinposed of periodate oxidation, Sodium borohydride reduction, Con A, and horseradish peroxidase reaction. Recently it was showin that these mucous cells secrete glycoproteins having GlcNAc alpha 1 to 4 Gal beta to R at nonreducing terminals of the carbohydrate moieties. Herein we describe the expression cloning of a cDNA encoding a human alpha l,4-N-acetylglucosaminyltransferase (aalpha 4 GnT), a key enzyme for the formation of GlcNAc lapha l to Gal beta 1 to R COS-1 cells were thus cotransfected with a stomach cDNA library and a leukosialin cDNA. Transfected COS-1 cells were screened by using monoclonal antibodies specific for GlcNAcalpha 1 to 4 Gal beta to R and coriched by fluorescence-activated cell sorting. Sibling selection of recovered plasmids resulted in a cDNA clone that directs the expression of GlcNAcalpah 1 to 4Gal beta to R, The deduced amino acid sequence predicts a type 11 membrane protein with 340 amino acids, showing no significant similarity with any other proteins. The alpha 4GnT gene is located at chromosome 3pl4.3, and its transcripts are expressed in the stomach and pancreas. An in vitro GIcNAc transferase assay by using a soluble cr4GnT revealed that alpha 1,4-linked GlcNAc residues are transferred most efficiently to core 2 branched O-glycans (Gal beta l- 4GlcNAc beta l to 6(Galbeta 1

机译：在分泌粘液的细胞中，胃腺粘液细胞，Brunnei腺，胰腺黏膜附属腺和表现出胃上皮化生的胰管是独特的，因为它们表达由高碘酸盐相悖的Con A染色鉴定的III类粘蛋白。氧化，硼氢化钠还原，Con A和辣根过氧化物酶反应。最近发现，这些粘液细胞在碳水化合物部分的非还原末端分泌具有GlcNAc alpha 1至4 Galβ1至R的糖蛋白。本文中，我们描述了编码人α1,4-N-乙酰氨基葡萄糖氨基转移酶（aalpha 4 GnT）的cDNA的表达克隆，从而将GlcNAc lapha 1形成为Gal beta 1到R COS-1细胞的关键酶与胃cDNA文库和白蛋白cDNA。通过使用对GlcNAcalpha 1至4 Gal beta至R特异的单克隆抗体筛选转染的COS-1细胞，并通过荧光激活细胞分选法使其富集。对回收质粒进行同胞选择会产生一个cDNA克隆，该克隆将GlcNAcalpah 1至4Gal beta的表达引导至R。推导的氨基酸序列预测具有340个氨基酸的11型膜蛋白，与任何其他蛋白均无显着相似性。 alpha 4GnT基因位于3pl4.3号染色体上，其转录本在胃和胰腺中表达。使用可溶性cr4GnT进行的体外GIcNAc转移酶测定显示，α1,4-连接的GlcNAc残基最有效地转移至核心2个分支的O-聚糖（Gal beta1-4GlcNAc beta1至6（Galbeta 1

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《Proceedings of the National Academy of Sciences of the United States of America》 |1999年第16期|8991-8996|共6页
作者
JUN NAKAYAMA; JIUNN-CHERN YEH; ANUP K.MISKRA;
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收录信息美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
原文格式 PDF
正文语种 eng
中图分类自然科学总论;
关键词
biochemistry; cell-type mucin; mucus-secreting cells; gastric metaplasia;

机译：生化;细胞型黏蛋白;黏液分泌细胞;胃化生;

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机译：在N-和O-聚糖中将硫酸盐转移至GalNAc beta 1-> 4GlcNAc beta 1-> R的两种截然不同的人N-乙酰半乳糖胺4-O-磺基转移酶的分子克隆和表达。
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机译：形成GlcNAcα1→4Galβ→R的人α14-N-乙酰氨基葡萄糖氨基转移酶的表达克隆该糖在胃腺黏液细胞型黏蛋白中特异性表达
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Expression cloning of a human alpha l,4-Nacetylglucosaminyltransfe that forms GlcNAc aphal l to 4 Gal beta to R, a glycan specificanlly expressed in the gastric gland mucous cell-type mucin

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