Structural determinants of sphingolipid recognition by commercially available anti-ceramide antibodies
Structural determinants of sphingolipid recognition by commercially available anti-ceramide antibodies
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DOI：
10.1194/jlr.m200241-jlr200
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发表时间：
2002-12-01
影响因子：
6.5
通讯作者：
Hannun, YA
中科院分区：
文献类型：
--
作者：
Cowart, LA;Szulc, Z;Hannun, YA
鞘脂介质神经酰胺参与细胞凋亡、分化、细胞因子反应和应激反应等细胞过程。实验证据表明，神经酰胺的细胞内位置可能是决定其最终细胞效应的关键因素。神经酰胺定位的一种方法是使用最近开发的抗神经酰胺抗体进行免疫细胞化学研究。现在有两种这样的商业制剂可供使用；我们试图比较和对比它们对神经酰胺和/或其他细胞脂质的特异性。通过使用脂质覆盖测定和多种鞘脂，我们能够描述这些试剂的特异性，从而描述其效用。我们的结果表明，其中一种抗神经酰胺制剂对神经酰胺和二氢神经酰胺具有相当的特异性，而另一种制剂则识别二氢神经酰胺、磷脂酰胆碱和鞘磷脂。此外，通过在类似的测定中使用化学修饰的神经酰胺，我们能够确定这两种试剂的脂质识别的一些结构决定因素。
The sphingolipid mediator ceramide is involved in cellular processes such as apoptosis, differentiation, responses to cytokines, and stress responses. Experimental evidence suggests that the intracellular location of ceramide may be a key factor in determining its ultimate cellular effects. One approach to ceramide localization is the use of recently developed anti-ceramide antibodies for immunocytochemical studies. Two such commercial preparations are now available; we sought to compare and contrast their specificity for ceramide and/or other cellular lipids. By using lipid overlay assays and a diverse panel of sphingolipids, we were able to delineate the specificity and thus, the utility of these reagents. Our results indicate that one of these anti-ceramide preparations is quite specific for ceramide and dihydroceramide, whereas the other preparation recognizes dihydroceramide, phosphatidylcholine, and sphingomyelin. Furthermore, through the use of chemically modified ceramides in similar assays, we were able to determine some structural determinants of lipid recognition by both of these reagents.