当前位置: X-MOL 学术bioRxiv. Biophys. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Single-molecule studies of conformational states and dynamics in the ABC importer OpuA
bioRxiv - Biophysics Pub Date : 2020-11-02 , DOI: 10.1101/2020.08.07.241463
Konstantinos Tassis , Ruslan Vietrov , Matthijs de Koning , Marijn de Boer , Giorgos Gouridis , Thorben Cordes

The current model of active transport via ABC importers is mostly based on structural, biochemical and genetic data. We here establish single-molecule Foerster-resonance energy transfer (smFRET) assays to monitor the conformational states and heterogeneity of the type-I ABC importer OpuA from Lactococcus lactis. Our studies include intradomain assays that elucidate conformational changes within the substrate-binding domain (SBD) OpuAC and interdomain assays between SBDs or transmembrane domains. Using the methodology, we studied ligand-binding mechanisms as well as ATP and glycine betaine dependences of conformational changes. Our study expands the scope of smFRET investigations towards a class of so far unstudied ABC importers, and paves the way for a full understanding of their transport cycle in the future.

中文翻译:

ABC进口商OpuA中的构象状态和动力学的单分子研究

当前通过ABC进口商进行主动运输的模型主要基于结构,生化和遗传数据。我们在这里建立单分子Foerster共振能量转移(smFRET)分析法,以监测来自乳酸乳球菌的I型ABC进口商OpuA的构象状态和异质性。我们的研究包括阐明底物结合域(SBD)OpuAC内构象变化的域内测定,以及SBD或跨膜结构域之间的域间测定。使用该方法,我们研究了配体结合机制以及构象变化的ATP和甘氨酸甜菜碱依赖性。我们的研究将smFRET研究的范围扩大到一类到目前为止尚未研究的ABC进口商,并为全面了解其未来的运输周期铺平了道路。
更新日期:2020-11-03
down
wechat
bug