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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Virulence of Leishmania major in macrophages and mice requires the gluconeogenic enzyme fructose-1,6-bisphosphatase
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Virulence of Leishmania major in macrophages and mice requires the gluconeogenic enzyme fructose-1,6-bisphosphatase

机译：利什曼原虫在巨噬细胞和小鼠中的毒力需要糖异生酶果糖-1,6-双磷酸酶

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摘要

Leishmania are protozoan parasites that replicate within mature phagolysosomes of mammalian macrophages. To define the biochemical composition of the phagosome and carbon source requirements of intracellular stages of L. major, we investigated the role and requirement for the gluconeogenic enzyme fructose-1,6-bisphosphatase (FBP). L. major FBP was constitutively expressed in both extracellular and intracellular stages and was primarily targeted to glycosomes, modified peroxisomes that also contain glycolytic enzymes. A L. major FBP-null mutant was unable to grow in the absence of hexose, and suspension in glycerol-containing medium resulted in rapid depletion of internal carbohydrate re- serves. L. major Delta fbp promastigotes were internalized by macrophages and differentiated into amastigotes but were unable to replicate in the macrophage phagolysosome. Similarly, the mutant persisted in mice but failed to generate normal lesions. The data suggest that Leishmania amastigotes reside in a glucose-poor phagosome and depend heavily on nonglucose carbon sources. Feeding experiments with [C-13]fatty acids showed that fatty acids are poor gluconeogenic substrates, indicating that amino acids are the major carbon source in vivo. The need for amino acids may have forced Leishmania spp. to adapt to life in the mature phagolysosome.
机译：利什曼原虫是在哺乳动物巨噬细胞的成熟吞噬体中复制的原生动物寄生虫。为了确定大叶利什曼原虫细胞阶段吞噬体的生物化学组成和碳源需求，我们调查了糖异生酶-1,6-双磷酸果糖（FBP）的作用和需求。 L. major FBP在细胞外和细胞内阶段均组成性表达，主要靶向糖体，修饰的过氧化物酶体，其中也含有糖酵解酶。在没有己糖的情况下，L。major FBP-null突变体无法生长，悬浮在含甘油的培养基中会导致内部碳水化合物储备的快速消耗。 L.主要Delta fbp前鞭毛体被巨噬细胞内化并分化成变形虫，但不能在巨噬细胞吞噬溶酶体中复制。同样，该突变体在小鼠中持续存在，但未能产生正常的病变。数据表明，利什曼原虫amastigotes居住在缺乏葡萄糖的吞噬体中，并严重依赖于非葡萄糖碳源。用[C-13]脂肪酸进料的实验表明，脂肪酸是不良的糖异生底物，表明氨基酸是体内主要的碳源。对氨基酸的需求可能迫使利什曼原虫属。适应生活中的成熟吞噬体。

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