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首页> 外文期刊>Infection and immunity >Characterization of Pseudomonas aeruginosa Exoenzyme S as a Bifunctional Enzyme in J774A.1 Macrophages
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Characterization of Pseudomonas aeruginosa Exoenzyme S as a Bifunctional Enzyme in J774A.1 Macrophages

机译：铜绿假单胞菌外切酶S作为J774A.1巨噬细胞中的双功能酶的表征

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摘要

Pseudomonas aeruginosa exoenzyme S (ExoS) is a type III secretion (TTS) effector, which includes both a GTPase-activating protein (GAP) activity toward the Rho family of low-molecular-weight G (LMWG) proteins and an ADP-ribosyltransferase (ADPRT) activity that targets LMWG proteins in the Ras, Rab, and Rho families. The coordinate function of both activities of ExoS in J774A.1 macrophages was assessed by using P. aeruginosa strains expressing and translocating wild-type ExoS or ExoS defective in GAP and/or ADPRT activity. Distinct and coordinated functions were identified for both domains. The GAP activity was required for the antiphagocytic effect of ExoS and was linked to interference of lamellopodium and membrane ruffle formation. Alternatively, the ADPRT activity of ExoS altered cellular adherence and morphology and was linked to effects on filopodium formation. The cellular mechanism of ExoS GAP activity included an inactivation of Rac1 function, as determined in p21-activated kinase 1-glutathione S-transferase (GST) pull-down assays. The ADPRT activity of ExoS targeted Ras and RalA but not Rab or Rho proteins, and Ral binding protein 1-GST pull-down assays identified an effect of ExoS ADPRT activity on RalA activation. The results from these studies confirm the bifunctional nature of ExoS activity within macrophages when translocated by TTS.
机译：铜绿假单胞菌外切酶S（ExoS）是一种III型分泌（TTS）效应物，包括针对低分子量G（LMWG）Rho家族的GTPase激活蛋白（GAP）活性。蛋白和针对Ras，Rab和Rho家族中LMWG蛋白的ADP-核糖基转移酶（ADPRT）活性。使用 P评估了J774A.1巨噬细胞中ExoS的两种活性的协调功​​能。表达和转移野生型ExoS或GAP和/或ADPRT活性有缺陷的ExoS的铜绿菌株。确定了两个领域的不同职能和协调职能。 GAP活性是ExoS的抗吞噬作用所必需的，并且与lamellopodium的干扰和膜褶皱的形成有关。另外，ExoS的ADPRT活性会改变细胞粘附和形态，并与对fi的形成有影响。 ExoS GAP活性的细胞机制包括Rac1功能的失活，这是通过p21激活的激酶1-谷胱甘肽 S -转移酶（GST）下拉测定法确定的。 ExoS靶向Ras和RalA而不是Rab或Rho蛋白的ADPRT活性，Ral结合蛋白1-GST下拉测定法确定了ExoS ADPRT活性对RalA激活的影响。这些研究的结果证实了被TTS转运时，巨噬细胞内ExoS活性的双重功能。

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