Damage of human renal glomerular endothelial cells (HRGECs) of the kidney represents the linchpin in the pathogenesis of the hemolytic uremic syndrome caused by Shiga toxins of enterohemorrhagic Escherichia coli (EHEC). We performed a comprehensive structural analysis of the Stx-receptor glycosphingolipids (GSLs) globotriaosylceramide (Gb3Cer, Galα4Galβ4Glcβ1Cer) and globotetraosylceramide (Gb4Cer, GalNAcβ3Galα4Galβ4Glcβ1Cer) and their distribution in lipid raft analog detergent-resistant membranes (DRMs) and nonDRMs prepared from primary HRGECs. Predominant receptor lipoforms were Gb3Cer and Gb4Cer with Cer (d18:1, C16:0), Cer (d18:1, C22:0) and Cer (d18:1, C24:1/C24:0). Stx-receptor GSLs co-distribute with sphingomyelin (SM) and cholesterol as well as flotillin-2 in DRMs, representing the liquid-ordered membrane phase and indicating lipid raft association. Lyso-phosphatidylcholine (lyso-PC) was identified as a nonDRM marker phospholipid of the liquid-disordered membrane phase. Exposure of primary HRGECs to the ceramide analogon d-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (D-PDMP) reduced total Gb3Cer and Gb4Cer content, roughly calculated from two biological replicates, down to half and quarter of its primordial content, respectively, but strengthened their prevalence and cholesterol preponderance in DRMs. At the same time, the distribution of PC, SM and lyso-PC to subcellular membrane fractions remained unaffected by D-PDMP treatment. Defining the GSL composition and precise microdomain structures of primary HRGECs may help to develop novel therapeutic options to combat life-threatening EHEC infections.

中文翻译：

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志贺毒素受体与脂筏在原代人肾小球内皮细胞中的共定位以及D-PDMP对糖鞘脂受体合成和分布的影响

肾脏的人肾小球内皮细胞（HRGEC）损伤代表由出血性大肠杆菌（EHEC）的志贺毒素引起的溶血性尿毒症综合征的发病机理中的关键。我们执行的STX-受体糖脂进行了全面的结构分析（的GSL）globotriaosylceramide（Gb3Cer，Galα4Galβ4Glcβ1Cer）和globotetraosylceramide（Gb4Cer，GalNAcβ3Galα4Galβ4Glcβ1Cer）及其分布脂筏由主要HRGEC制备的类似洗涤剂抗性膜（DRM）和非DRM。主要的受体脂质形式是Gb3Cer和Gb4Cer和Cer（d18：1，C16：0），Cer（d18：1，C22：0）和Cer（d18：1，C24：1 / C24：0）。Stx受体GSL与DRM中的鞘磷脂（SM）和胆固醇以及flotillin-2共分布，代表液序膜相并指示脂质筏缔合。溶血磷脂酰胆碱（lyso-PC）被鉴定为液体紊乱膜相的nonDRM标记磷脂。初级HRGECs到神经酰胺analogon的曝光d -苏式-1-苯基-2-癸酰氨基-3-吗啉代-1-丙醇（d-PDMP）降低了总的Gb3Cer和Gb4Cer含量（从两个生物学重复中粗略计算），分别降至其原始含量的一半和四分之一，但是却增强​​了它们在DRM中的患病率和胆固醇优势。同时，PC，SM和溶血-PC在亚细胞膜部分的分布不受D-PDMP处理的影响。定义主要HRGEC的GSL组成和精确的微结构可能有助于开发新的治疗选择，以对抗威胁生命的EHEC感染。