cDNA cloning and expression of a novel human UDP-N-acetyl-alpha-D-galactosamine - Polypeptide N-acetylgalactosaminyltransferase, GalNAc-T3
cDNA cloning and expression of a novel human UDP-N-acetyl-alpha-D-galactosamine - Polypeptide N-acetylgalactosaminyltransferase, GalNAc-T3
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DOI：
10.1074/jbc.271.29.17006
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发表时间：
1996-07-19
影响因子：
4.8
通讯作者：
Clausen, H
中科院分区：
文献类型：
--
作者：
Bennett, EP;Hassan, H;Clausen, H
丝氨酸和苏氨酸残基在粘蛋白型0-连接蛋白糖基化期间的糖基化通过UDP-GalNAc：多肽N-乙酰半乳糖胺转移酶（GalNAc-转移酶）家族进行。以前的两个成员，GalNAc-T1和-T2，已被分离和基因克隆和表征，在这里，我们报告的cDNA克隆和表达的一个新的GalNAc-转移酶称为GalNAc-T3。基于半乳糖NAc转移酶基序的鉴定，分离并克隆了该基因该cDNA序列具有633个氨基酸的编码区，指示具有II型结构域结构的72.5kDa的蛋白质，与GalNAc-T1和GalNAc-T2的总氨基酸序列相似性约为45%;在GalNAc-T1和-T2之间共享的12个半胱氨酸残基也存在于GalNAcT 3中，利用杆状病毒载体在昆虫细胞中将GalNAc-T3表达为不含疏水跨膜结构域的可溶性蛋白，并且表达的GalNAc-转移酶活性显示出与先前报道的GalNAc-T1和GalNAc-T2不同的底物特异性，人体器官的北方分析揭示了GalNAc-T3的非常有限的表达模式。
The glycosylation of serine and threonine residues during mucin-type O-linked protein glycosylation is carried out by a family of UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferases (GalNAc-transferase). Previously two members, GalNAc-T1 and -T2, have been isolated and the genes cloned and characterized, Here we report the cDNA cloning and expression of a novel GalNAc-transferase termed GalNAc-T3. The gene was isolated and cloned based on the identification of a GalNAc-transferase motif (61 amino acids) that is shared between GalNAc-T1 and -T2 as well as a homologous Caenorhabditis elegans gene, The cDNA sequence has a 633-amino acid coding region indicating a protein of 72.5 kDa with a type II domain structure, The overall amino acid sequence similarity with GalNAc-T1 and -T2 is approximately 45%; 12 cysteine residues that are shared between GalNAc-T1 and -T2 are also found in GalNAcT3, GalNAc-T3 was expressed as a soluble protein without the hydrophobic transmembrane domain in insect cells using a Baculo-virus vector, and the expressed GalNAc-transferase activity showed substrate specificity different from that previously reported for GalNAc-T1 and -T2, Northern analysis of human organs revealed a very restricted expression pattern of GalNAc-T3.