Involvement of murine β-1,4-galactosyltransferase V in lactosylceramide biosynthesis.
Involvement of murine β-1,4-galactosyltransferase V in lactosylceramide biosynthesis.
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DOI：
10.1007/s10719-010-9313-2
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发表时间：
2010-10
影响因子：
3
通讯作者：
Furukawa, Kiyoshi
中科院分区：
文献类型：
--
作者：
Kumagai, Tadahiro;Sato, Takeshi;Natsuka, Shunji;Kobayashi, Yukito;Zhou, Dapeng;Shinkai, Tadashi;Hayakawa, Satoru;Furukawa, Kiyoshi
体外研究表明，人β-1，4-半乳糖基转移酶（β-1，4-GalT）V参与N-聚糖、O-聚糖和乳糖神经酰胺（Lac-Cer）的生物合成。为了确定其底物特异性，使用来自β-1，4-GalT V（B4 galt 5）-突变小鼠的小鼠胚胎成纤维细胞（MEF）分析酶活性及其产物。对β-1，4-GalT I-VI基因的表达水平的分析显示，与B4 galt 5 +/+-来源的MEF细胞相比，B4 galt 5 +/−和B4 galt 5 −/−来源的MEF细胞中β-1，4-GalT V基因的表达为一半和零，而不改变其他β-1，4-GalT基因的表达水平。这些MEF细胞的生长没有明显差异。当确定β-1，4-GalT对GlcNAcβ-S-pNP的活性时，在B4 galt 5 +/+-、B4 galt 5 +/−-和B4 galt 5 −/−-衍生的MEF细胞中，其比活性没有显著差异。在B4 galt 5 +/+-、B4 galt 5 +/−-和B4 galt 5 −/−-来源的MEF细胞中，N-聚糖和凝集素与膜糖蛋白的结合的结构和量没有显著差异。然而，当细胞匀浆在UDP-[3 H]Gal存在下与葡糖基神经酰胺一起孵育时，B4 galt 5 +/−和B4 galt 5 −/−来源的MEF细胞中的Lac-Cer合酶活性分别降低至B4 galt 5 +/+来源的MEF细胞的41%和11%。与此一致，与B4 galt 5 +/+来源的MEF细胞相比，B4 galt 5 −/−来源的MEF细胞中Lac-Cer及其衍生物GM 3的量显著减少。这些结果表明鼠β-1，4-GalT V参与Lac-Cer生物合成。
Human β-1,4-galactosyltransferase (β-1,4-GalT) V was shown to be involved in the biosynthesis of N-glycans, O-glycans and lactosylceramide (Lac-Cer) by in vitro studies. To determine its substrate specificity, enzymatic activity and its products were analyzed using mouse embryonic fibroblast (MEF) cells from β-1,4-GalT V (B4galt5)-mutant mice. Analysis of expression levels of the β-1,4-GalT I-VI genes revealed that the expression of the β-1,4-GalT V gene in B4galt5+/−- and B4galt5−/−-derived MEF cells are a half and null when compared to that of B4galt5+/+-derived MEF cells without altering the expression levels of other β-1,4-GalT genes. These MEF cells showed no apparent difference in their growth. When β-1,4-GalT activities were determined towards GlcNAcβ-S-pNP, no significant difference in its specific activity was obtained among B4galt5+/+-, B4galt5+/−- and B4galt5−/−-derived MEF cells. No significant differences were obtained in structures and amounts of N-glycans and lectin bindings to membrane glycoproteins among B4galt5+/+-, B4galt5+/−- and B4galt5−/−-derived MEF cells. However, when cell homogenates were incubated with glucosylcer-amide in the presence of UDP-[3H]Gal, Lac-Cer synthase activity in B4galt5+/−- and B4galt5−/−-derived MEF cells decreased to 41% and 11% of that of B4galt5+/+-derived MEF cells. Consistent with this, amounts of Lac-Cer and its derivative GM3 in B4galt5−/−-derived MEF cells decreased remarkably when compared with those of B4galt5+/+ derived MEF cells. These results indicate that murine β-1,4-GalT V is involved in Lac-Cer biosynthesis.
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影响因子：
4.4
作者：
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Biellmann F;Hülsmeier AJ;Zhou D;Cinelli P;Hennet T
通讯作者：
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