期刊
IUBMB LIFE
卷 74, 期 7, 页码 715-722出版社
WILEY
DOI: 10.1002/iub.2618
关键词
CyaY; desulfurase; enzymatic activity; frataxin; Friedreich's ataxia; iron-sulfur cluster biogenesis
资金
- University of Turin [ADIS RILO 18 01]
- Alzheimer's Research, UK
- Medical Research Council
- Dementia Research Institute [RE1 3556]
- Institut Pasteur
This study proposes a new approach for studying iron-sulfur cluster biogenesis using genetically engineered bacterial strains and metabolomics. The ex vivo method closely reproduces the in vitro results while retaining the complexity of the system.
Iron-sulfur clusters are prosthetic groups that are assembled on their acceptor proteins through a complex machine centered on a desulfurase enzyme and a transient scaffold protein. Studies to establish the mechanism of cluster formation have so far used either in vitro or in vivo methods, which have often resulted in contrasting or non-comparable results. We suggest, here, an alternative approach to study the enzymatic reaction, that is based on the combination of genetically engineered bacterial strains depleted of specific components, and the detection of the enzymatic kinetics in cellular extracts through metabolomics. Our data prove that this ex vivo approach closely reproduces the in vitro results while retaining the full complexity of the system. We demonstrate that co-presence of bacterial frataxin and iron is necessary to observe an inhibitory effect of the enzymatic activity of bacterial frataxin. Our approach provides a new powerful tool for the study of iron-sulfur cluster biogenesis.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据