Purification and Analysis of Effector Glucosyltransferase Lgt1 from Legionella pneumophila.

Purification and Analysis of Effector Glucosyltransferase Lgt1 from Legionella pneumophila.
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嗜肺军团菌效应葡萄糖基转移酶 Lgt1 的纯化和分析

DOI:
10.1007/978-1-4939-9048-1_18
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发表时间:
1921
影响因子:
--
通讯作者:
Belyi Y
Belyi Y
中科院分区:
--
文献类型:
--
作者:
Levanova N;Tabakova I;Jank T;Belyi Y

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嗜肺军团菌是一种兼性细胞内病原体,可引起军团菌病,一种严重的人类肺部疾病。这种细菌使用4 b型分泌系统将各种效应蛋白递送到真核靶细胞的细胞质中。其中之一是葡萄糖基转移酶Lgt 1。该效应器通过单-O-葡糖基化修饰真核细胞延伸因子1A(eEF 1A)中的丝氨酸-53。eEF 1A的修饰导致蛋白质合成的抑制和真核细胞的死亡,这些过程被认为有助于军团菌感染。在这里,我们描述了一个协议的葡萄糖基转移酶Lgt 1从mL的分离。随后使用14 C-UDP-葡萄糖放射自显影术测定其酶活性。
Legionella pneumophilais a facultative intracellular pathogen responsible for legionellosis, a severe lung disease in humans. This bacterium uses a type 4b secretion system to deliver various effector proteins into the cytoplasm of a eukaryotic target cell. Among those is the glucosyltransferase Lgt1. This effector modifies serine-53 in eukaryotic elongation factor 1A (eEF1A) by mono-O-glucosylation. Modification of eEF1A results in inhibition of protein synthesis and death of the eukaryotic cell, processes which are thought to contribute toLegionellainfection. Here we describe a protocol for isolation of the glucosyltransferase Lgt1 fromL. pneumophilaculture followed by assaying its enzymatic activity using14C-UDP-glucose autoradiography.
酵母延伸因子复合物 eEF1A-eEF1B α 的结晶。
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