HdeA在细菌外膜囊泡环境下的原位NMR研究
收稿日期: 2023-05-16
网络出版日期: 2023-06-30
基金资助
国家自然科学基金资助项目(22174151);国家自然科学基金资助项目(21904138);国家自然科学基金资助项目(21991080);国家自然科学基金资助项目(22078280);国家重点基础研究发展计划（973计划）资助项目(2018YFE0202301);国家重点基础研究发展计划（973计划）资助项目(2018YFE0202300)
In situ Investigation of HdeA in Bacterial Outer Membrane Vesicles Using NMR Spectroscopy
Received date: 2023-05-16
Online published: 2023-06-30
HdeA是一种定位于细菌周质的分子伴侣，在维持蛋白质稳态中起着重要的作用．以往对HdeA的研究主要是在体外条件下进行，限制了人们对HdeA在天然环境下发挥作用机制的理解．细菌外膜囊泡是细菌自发分泌到胞外环境的外膜囊泡，其内容物与周质环境相似．本研究将HdeA富集到细菌外膜囊泡（OMVs）中，通过核磁共振波谱研究HdeA在OMVs中的构象变化．结果表明，HdeA在其原位环境中表现出酸依赖性的构象变化．在低pH条件下HdeA主要通过S15、W16、T17、S27、T32、E36、G54、T57、C66、Q71、F74及D83等残基启动其分子伴侣功能．此外本研究也为原位研究其它周质分子伴侣的功能提供了新方法．
王欢 , 陶志清 , 姜国胜 , 张许 , 王冠 , 禾立春 , 刘买利 . HdeA在细菌外膜囊泡环境下的原位NMR研究[J]. 波谱学杂志, 2024 , 41(1) : 1 -8 . DOI: 10.11938/cjmr20233069
HdeA, a molecular chaperone localized in the bacterial periplasm, plays a vital role in maintaining protein homeostasis. Previous studies of HdeA were mainly carried out in in vitro conditions, limiting our understanding of the mechanism of HdeA in its native environment. Outer membrane vesicles of bacteria are extracellular vesicles released by cells into the extracellular milieu in a controlled manner, with contents almost identical to the periplasmic environment. Thus, by enriching HdeA into the bacterial outer membrane vesicles (OMVs), we investigated the conformational changes of HdeA within OMVs via nuclear magnetic resonance (NMR) spectroscopy. The results reveal that HdeA exhibits an acid-dependent conformational change in its native environment. The high-resolution spectrum of HdeA in OMVs indicates that under low pH conditions the function of HdeA is activated through residues S15, W16, T17, S27, T32, E36, G54, T57, C66, Q71, F74 and D83. Moreover, the NMR measurement of HdeA within OMVs provides a promising way for in situ investigation of mechanisms of other periplasmic molecular chaperones via NMR spectroscopy.
Key words： in situ NMR; HdeA; outer membrane vesicles; chaperone
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