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Engineering a Cl--Modulated Light-Driven Na+ Pump
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DOI：10.1002/chem.202302543.png)
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摘要
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Microbial Na+-pumping rhodopsin (NaR) is a promising optogenetic tool due to its unique ability to transport Na+. Like most rhodopsin-based tools, NaR is limited to light-based control. In this study, our objective was to develop a novel mode of modulation for NaR beyond light control. By introducing a potential Cl- binding site near the putative Na+ release cavity, we engineered Nonlabens dokdonensis rhodopsin 2 (NdR2) to be modulated by Cl-, an essential chemical in organisms. The engineered NdR2 demonstrated an approximately two-fold increase in Na+ pump activity in the presence of 100 mM Cl- compared to Cl--free solution. Increasing Cl- concentration decreased the lifetimes of the M and O intermediates accordingly. The analysis of competitive ion uptake suggested the bound Cl- may increase the Na+ affinity and selectivity. This chemical modulation allows for more diverse and precise control over cellular processes, advancing the development of next-generation optogenetic tools. Notably, our Cl--modulated NdR2 establishes an innovative mechanism for linking Cl- to Na+-related processes, with potential applications in optogenetic therapies for related diseases.
Keyword:
chloride modulation
kinetics
membrane proteins
sodium pumps
期刊
C
IF:
3.7
论文数:
3.9W
被引数:
9.6W
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