IMPROVED INHIBITORS OF GLUCOSYLCERAMIDE SYNTHASE
IMPROVED INHIBITORS OF GLUCOSYLCERAMIDE SYNTHASE
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DOI：
10.1093/oxfordjournals.jbchem.a123736
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发表时间：
1992-02-01
影响因子：
2.7
通讯作者：
RADIN, NS
中科院分区：
文献类型：
--
作者：
ABE, A;INOKUCHI, J;RADIN, NS
据报道，葡萄糖神经酰胺(GlcCer)合成酶的抑制剂1-phenyl-2-decanoylamino-3-morpholino-1-propanol(PDMP)会耗尽细胞和小鼠的鞘糖脂。该抑制剂已被证明对阐明该脂质家族的许多功能很有用[由Radin，N.S.和Inokuchi，J.(1991)Trends Glycoci审查。甘草甜素。3、200-213]。在本研究中，我们合成了具有不同酰基链的PDMP同系物(C6-C18)，并比较了它们在体外对GlcCer合成酶的抑制作用和对培养的MDCK(Madin-Darby犬肾)细胞GlcCer、蛋白质和DNA合成的抑制作用。利用MDCK匀浆和小鼠脑和肝微粒体，我们发现C6化合物活性相对较低，较长链的化合物在抑制活性方面没有太大差异。然而，使用完整的MDCK细胞表明，较长链同系物在抑制GlcCer合成、细胞生长和[H-3]胸腺嘧啶核苷掺入方面要有效得多。对两个放射性同系物的测试表明，具有较长酰链的抑制剂被MDCK细胞更有效地摄取，这种差异解释了这种同系物在完整细胞中的更大有效性。无论是用非离子洗涤剂还是用牛血清白蛋白增溶，抑制剂都是有效的。DNA合成的减少程度与细胞内葡萄糖神经酰胺的减少不成正比，可能是因为DNA合成只需要较低水平的糖脂。
An inhibitor of glucosylceramide (GlcCer) synthase, 1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP), has been reported to deplete cells and mice of their glucosphingolipids. This inhibitor has proved useful for the elucidation of the many functions of this lipid family [reviewed by Radin, N.S. & Inokuchi, J. (1991) Trends Glycosci. Glycotechnol. 3, 200-213]. In the present study, we have synthesized homologs of PDMP having different acyl chains (C6-C18) and compared their effectiveness for the inhibition of GlcCer synthase in vitro and their inhibition of GlcCer, protein, and DNA synthesis in cultured MDCK (Madin-Darby canine kidney) cells. Using MDCK homogenates and mouse brain and liver microsomes, we found that the C6 compound was relatively inactive and that the longer chain compounds did not differ much in inhibitory power. However, the use of intact MDCK cells showed that the longer chain homologs were much more effective in inhibiting GlcCer synthesis, cell growth, and incorporation of [H-3]thymidine. Tests with two radioactive homologs showed that the inhibitor with a longer acyl chain was taken up much more effectively by MDCK cells and that this difference explains the much greater effectiveness of this homolog in intact cells. The inhibitors were effective when solubilized either with a nonionic detergent or with bovine serum albumin. The extent of decrease in DNA synthesis was not directly proportional to the decrease in cellular glucosylceramide, possibly because only a low level of the glycolipid is needed for DNA synthesis.