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Phenylmethylsulfonyl fluoride (PMSF)

Company: NACALAI TESQUE
Catalog#: 27327-94
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Self-organization Assay for Min Proteins of Escherichia coli in Micro-droplets Covered with Lipids
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Date:
2020-03-20
[Abstract]  The Min system determines the cell division plane of bacteria. As a cue of spatiotemporal regulation, the Min system uses wave propagation of MinD protein (Min wave). Therefore, the reconstitution of the Min wave in cell-sized closed space will lead to the creation of artificial cells capable of cell division. The Min waves emerge via coupling between the reactions among MinD, MinE, and ATP and the differences in diffusion rate on the cell membrane and in the cytoplasm. Because Min waves appear only under the balanced condition of the reaction-diffusion coupling, special attentions are needed towards several technical points for the reconstitution of Min waves in artificial cells. This protocol describes a technical method for stably generating Min waves in artificial cells. [摘要]  [摘要 ] Min系统确定细菌的细胞分裂平面。作为时空调节的提示,Min系统使用MinD 蛋白的波传播(Min wave)。因此,Min波在细胞大小的封闭空间中的重构将导致能够分裂细胞的人造细胞的产生。闵波出现经由耦合之间反应小号中MinD的,的MinE ,和ATP 和所述differenc ES 在细胞膜上的扩散速度和在细胞质中。因为最小波仅在反应扩散耦合的平衡条件下出现, 特别关注,需要对几个技术要点为闽波在人造细胞重建。该协议描述了一种在人造细胞中稳定产生Min波的技术方法。

[背景 ] 敏系统,它决定了细胞分ER 对称细胞分裂,是在细菌细胞内的组织系统的最显着的例子之一(Rothfield 等人,2005;和罗利特马戈林,2013年)。敏系统使用图案形成在细胞内的时间依赖性蛋白梯度的公知的作为敏波(宽松等人,2008; Halatek和Frey,2012;邦尼等人,2013; Zieske 。等人,2016 ; Kohyama 。等人, 2019 )。Min波是由两种蛋白MinD 和MinE 的反应扩散耦合产生的。通过与ATP结合,MinD 形成二聚体并附着在膜上。的MinE 被招募到的ATP MinD的和诱导ATP酶的活性MinD的。通过MinE ,ATP- MinD 变为ADP- MinD ,并从膜上脱离。ADP- MinD的被转换回ATP- ...

Isolation of the Dot/Icm Type IV Secretion System Core Complex from Legionella pneumophila for Negative Stain Electron Microscopy Studies
Author:
Date:
2017-04-20
[Abstract]  Legionella possesses a pivotal secretion machinery to deliver virulence factors to eukaryotic host cells. In this protocol, we describe the procedure for isolation of the native core complex of the Dot/Icm type IV secretion system from L. pneumophila aiming to perform biochemical and transmission electron microscopy analyses. [摘要]  军团菌具有关键的分泌机制,以将毒力因子递送至真核宿主细胞。在本协议中,我们描述了从L / L分离Dot / Icm IV型分泌系统的天然核心复合物的步骤。肺炎支原体旨在进行生物化学和透射电子显微镜分析。

嗜肺军团菌是革兰氏阴性细菌病原体,其导致被称为退伍军人病的肺部感染(Fields等人,2002)。 L。嗜肺杆菌利用由 dot / cm 基因编码的IV型分泌系统(T4SS)将大约300种细菌蛋白转运到其真核宿主细胞质中以劫持细胞过程(Hubber和Roy, 2010)。由超过20种蛋白组成,T4SS是建立在细菌内膜和外膜上的纳米机器(Nagai和Kubori 2011; Kubori和Nagai 2016)。 Dot / Icm T4SS的核心复合物是系统的生物化学稳定部分,并形成桥接内膜和外膜的输送导管(Kubori等人,2014)。核心复合物由至少五种蛋白质组成;三种外膜相关蛋白,DotC,DotD和DotH,以及两种内膜蛋白DotF和DotG(Vincent等人,2006)。基于来自鼠伤寒沙门氏菌的另一种细菌纳米机械的III型分泌系统的生物化学分离方法(Kubori等人,1998; Marlovits等人, ...

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