首页> 外文OA文献 >Isolation and identification of Enterococcus faecalis membrane proteins using membrane shaving, 1D SDS/PAGE, and mass spectrometry
【2h】

Isolation and identification of Enterococcus faecalis membrane proteins using membrane shaving, 1D SDS/PAGE, and mass spectrometry

机译:使用膜剃刮,1D sDs / paGE和质谱分离和鉴定粪肠球菌膜蛋白

代理获取
本网站仅为用户提供外文OA文献查询和代理获取服务,本网站没有原文。下单后我们将采用程序或人工为您竭诚获取高质量的原文,但由于OA文献来源多样且变更频繁,仍可能出现获取不到、文献不完整或与标题不符等情况,如果获取不到我们将提供退款服务。请知悉。

摘要

Enterococcus faecalis is a significant nosocomial pathogen, which is able to survive in diverse environments and resist killing with antimicrobial therapies. The expression of cell membrane proteins play an important role in how bacteria respond to environmental stress. As such, the capacity to identify and study membrane protein expression is critical to our understanding of how specific proteins influence bacterial survival. Here, we describe a combined approach to identify membrane proteins of E. faecalis ATCC V583 using membranes fractionated by either 1D SDS/PAGE or membrane shaving, coupled with LC-ESI mass spectrometry. We identified 222 membrane-associated proteins, which represent approximately 24% of the predicted membrane-associated proteome: 170 were isolated using 1D SDS/PAGE and 68 with membrane shaving, with 36 proteins being common to both the techniques. Of the proteins identified by membrane shaving, 97% were membrane-associated with the majority being integral membrane proteins (89%). Most of the proteins identified with known physiology are involved with transportation across the membrane. The combined 1D SDS/PAGE and membrane shaving approach has produced the greatest number of membrane proteins identified from E. faecalis to date. These protocols will aid future researchers investigating changes in the membrane proteome of E. faecalis by improving our understanding of how E. faecalis adapts and responds to its environment.
机译:粪肠球菌是一种重要的医院病原体,能够在多种环境中生存并能通过抗菌疗法抵抗杀伤。细胞膜蛋白的表达在细菌对环境压力的反应中起着重要作用。因此,识别和研究膜蛋白表达的能力对于我们了解特定蛋白如何影响细菌存活至关重要。在这里,我们描述了一种结合方法,通过一维SDS / PAGE或膜剃刮分离的膜,结合LC-ESI质谱法,鉴定粪肠球菌ATCC V583的膜蛋白。我们鉴定了222种与膜相关的蛋白质,约占预测的与膜相关的蛋白质组的24%:使用1D SDS / PAGE分离了170种膜,用剃刮膜分离了68种,两种技术共有36种蛋白质。在通过剃刮膜鉴定出的蛋白质中,有97%与膜相关,而大多数是完整的膜蛋白质(占89%)。以已知生理学鉴定的大多数蛋白质都与跨膜运输有关。迄今为止,结合1D SDS / PAGE和膜剃须方法产生的粪便肠杆菌中鉴定出的膜蛋白数量最多。这些协议将通过增进我们对粪肠球菌如何适应和响应其环境的理解,来帮助未来的研究人员调查粪肠球菌的膜蛋白质组的变化。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
代理获取

客服邮箱:kefu@zhangqiaokeyan.com

京公网安备:11010802029741号 ICP备案号:京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有
  • 客服微信

  • 服务号