Luo Yi, Haltiwanger Robert S
Department of Biochemistry and Cell Biology, Institute for Cell and Developmental Biology, State University of New York at Stony Brook, Stony Brook, New York 11794-5215, USA.
J Biol Chem. 2005 Mar 25;280(12):11289-94. doi: 10.1074/jbc.M414574200. Epub 2005 Jan 14.
LADII (leukocyte adhesion deficiency type II)/CDGIIc (congenital disorder of glycosylation type IIc) is a rare autosomal recessive disease characterized by leukocyte adhesion deficiency as well as severe neurological and developmental abnormalities. It is caused by mutations in the Golgi GDP-fucose transporter, resulting in a reduction of fucosylated antigens on the cell surface. A recent study using fibroblasts from LADII/CDGIIc patients suggested that although terminal fucosylation of N-glycans is reduced severely, protein O-fucosylation is generally unaffected (Sturla, L., Rampal, R., Haltiwanger, R. S., Fruscione, F., Etzioni, A., and Tonetti, M. (2003) J. Biol. Chem. 278, 26727-26733). A potential explanation for this phenomenon is that enzymes adding O-fucose to proteins localize to cell organelles other than the Golgi apparatus. In this study, we investigated the subcellular localization of protein O-fucosyltransferase 1 (O-FucT-1), which is responsible for adding O-fucose to epidermal growth factor-like repeats. Our analysis reveals that, unlike all other known fucosyltransferases, O-FucT-1 is a soluble protein that localizes to the endoplasmic reticulum (ER). In addition, it appears that O-FucT-1 is retained in the ER by a KDEL-like sequence at its C terminus. Our results also suggest that enzymatic addition of O-fucose to proteins occurs in the ER, suggesting that a novel, ER-localized GDP-fucose transporter may exist. The fact that O-FucT-1 recognizes properly folded epidermal growth factor-like repeats, together with this unique localization, suggests that it may play a role in quality control.
II型白细胞黏附缺陷症(LADII)/IIc型糖基化先天性疾病(CDGIIc)是一种罕见的常染色体隐性疾病,其特征为白细胞黏附缺陷以及严重的神经和发育异常。它是由高尔基体GDP-岩藻糖转运蛋白的突变引起的,导致细胞表面岩藻糖基化抗原减少。最近一项使用LADII/CDGIIc患者成纤维细胞的研究表明,尽管N-聚糖的末端岩藻糖基化严重减少,但蛋白质O-岩藻糖基化通常不受影响(斯特拉,L.,兰帕尔,R.,哈尔蒂旺格,R.S.,弗鲁西奥内,F.,埃齐奥尼,A.,和托内蒂,M.(2003年)《生物化学杂志》278卷,26727 - 26733页)。对此现象的一种可能解释是,给蛋白质添加O-岩藻糖的酶定位于高尔基体以外的细胞器。在本研究中,我们调查了负责给表皮生长因子样重复序列添加O-岩藻糖的蛋白质O-岩藻糖基转移酶1(O-FucT-1)的亚细胞定位。我们的分析显示,与所有其他已知的岩藻糖基转移酶不同,O-FucT-1是一种定位于内质网(ER)的可溶性蛋白质。此外,它似乎通过其C末端的类KDEL序列保留在内质网中。我们的结果还表明,蛋白质的O-岩藻糖酶促添加发生在内质网中,这表明可能存在一种新型的、定位于内质网的GDP-岩藻糖转运蛋白。O-FucT-1能够识别正确折叠的表皮生长因子样重复序列,再加上这种独特的定位,表明它可能在质量控制中发挥作用。