CELL-FREE SYNTHESIS AND FUNCTIONAL CHARACTERIZATION OF SPHINGOLIPID SYNTHASES

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 布氏锥虫基因组有四个高度相似的基因编码鞘脂合成酶(TbSLS1-4)。TbSLS是多表膜蛋白，对这种人类原虫致病血流阶段的生存是必不可少的，因此，可以被认为是潜在的药物靶点。TbSLS4是一个双功能的鞘磷脂/乙醇胺磷酸神经酰胺合成酶，而其他的功能尚未确定。利用最近描述的脂质体补充的无细胞合成系统，消除了背景细胞活动的并发症，我们现在明确地定义了整个基因家族的酶特异性。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The Trypanosoma brucei genome has four highly similar genes encoding sphingolipid synthases (TbSLS1-4). TbSLSs are polytopic membrane proteins that are essential for viability of the pathogenic bloodstream stage of this human protozoan parasite and, consequently, can be considered as potential drug targets. TbSLS4 was shown previously to be a bifunctional sphingomyelin/ethanolamine phosphorylceramide synthase, whereas functions of the others were not characterized. Using a recently described liposome-supplemented cell-free synthesis system, which eliminates complications from background cellular activities, we now unambiguously define the enzymatic specificity of the entire gene family.