Hansson G C, Simons K, van Meer G
EMBO J. 1986 Mar;5(3):483-9. doi: 10.1002/j.1460-2075.1986.tb04237.x.
The glycosphingolipids (GSLs) of two sublines of Madin-Darby canine kidney (MDCK) cells, an epithelial cell line, were characterized by t.l.c., antibody overlay and mass spectrometry. The major characteristic which distinguishes the two MDCK cell strains is their trans-epithelial electrical resistance which is typically of the order of 3000 ohm.cm2 for strain I and 100 ohm.cm2 for strain II cells. Strain I and II cells were equally rich in glycolipids, the cellular GSL/phospholipid ratio being 0.04. However, while the phospholipid patterns were identical, the GSLs showed striking differences, and each cell strain expressed appreciable amounts of GSLs that were not found in the other strain. Both cell types possessed neutral GSLs with one, two or three carbohydrate moieties. The monoglycosylceramide accounted for 50% of the total GSLs in each strain. However, while in strain I cells over 90% of this monoglycosylceramide was monoglucosylceramide, in strain II cells over 90% consisted of monogalactosylceramide. In addition, MDCK strain II cells selectively expressed GSLs belonging to the globo series (26% of its neutral GSLs), including globoside and Forssman antigen, a globoside derivative. MDCK strain I cells, on the other hand, expressed another series of GSLs with 4-7 carbohydrate moieties characterized by the common sequence Hex-HexNAc-Hex-Hex-Cer. The presence of two fucosylated GSLs in these series was established. Both MDCK strain I and II cells contained negatively charged GSLs, the major component of which was the ganglioside GM3. MDCK strain II cells in addition expressed sulfatide, the sulfated derivative of galactosylceramide.(ABSTRACT TRUNCATED AT 250 WORDS)
采用薄层层析、抗体覆盖法及质谱分析法对上皮细胞系——马-达二氏犬肾(MDCK)细胞的两个亚系的糖鞘脂(GSL)进行了表征。区分这两种MDCK细胞株的主要特征是它们的跨上皮电阻,I株细胞的跨上皮电阻通常约为3000欧姆·平方厘米,II株细胞的则为100欧姆·平方厘米。I株和II株细胞的糖脂含量相当,细胞GSL/磷脂比为0.04。然而,虽然磷脂模式相同,但GSL却表现出显著差异,且每个细胞株都表达了大量在另一株中未发现的GSL。两种细胞类型都拥有带有一个、两个或三个碳水化合物部分的中性GSL。单糖基神经酰胺占各株总GSL的50%。然而,I株细胞中超过90%的这种单糖基神经酰胺是单葡萄糖神经酰胺,而在II株细胞中超过90%是单半乳糖神经酰胺。此外,MDCK II株细胞选择性表达属于球系列的GSL(占其中性GSL的26%),包括红细胞糖苷脂和福斯曼抗原(一种红细胞糖苷脂衍生物)。另一方面,MDCK I株细胞表达了另一系列带有4 - 7个碳水化合物部分的GSL,其特征序列为己糖-己糖胺-己糖-己糖-神经酰胺。已确定这些系列中存在两种岩藻糖基化的GSL。MDCK I株和II株细胞都含有带负电荷的GSL,其主要成分是神经节苷脂GM3。MDCK II株细胞还表达了硫苷脂,即半乳糖神经酰胺的硫酸化衍生物。(摘要截选至250词)