Collaboration leads to breakthroughs
The research group’s complementary expertise was key to making the discovery. Through electrophysiology and structural studies, the Rockefeller team was able to guide the placement of single-molecule probes. St. Jude team. By deploying single-molecule fluorescence resonance energy transfer (smFRET), St. Jude The team was able to provide new insight into the moving parts of the CFTR mechanism. By integrating cryo-electron microscopy, electrophysiology and smFRET, the research group was able to draw the connections needed to better understand how his CFTR works.
“There is potential here to help patients with cystic fibrosis by learning about the structure and behavior of CFTR,” said first author Jesper Levring of The Rockefeller University. “Using single-channel electrophysiology and these methods he called smFRET, he looked at these molecules one by one, correlating the function of the channel with conformational changes, and drawing on the underlying structural biology.” can be associated.”
What the researchers found is that CFTR exhibits a hierarchical gating mechanism. The two nucleotide-binding domains of CFTR dimerize (bind) before the channel opens. A conformational change within the dimerization channel associated with ATP hydrolysis, a reaction in which energy is released, regulates chloride conductance. The importance of this mechanistic insight is further demonstrated by the finding that the potentiators Ivacaftor and GLPG1837 enhance channel activity by increasing pore opening while the nucleotide-binding domain is dimerized. became. Mutations that cause cystic fibrosis can reduce the efficiency of dimerization. These insights will help in the search for more effective clinical therapies.
“What I find most satisfying about this study is that it answers a long-debated question in the field about how CFTR works,” said Chen, co-corresponding author of the study. “Even if we have good data, we won’t get the answer because each individual method has its own limitations. Combining the approaches will give us a unified mechanism that will give us insight into how this molecule works.” This understanding will allow us to test how mutations and drugs affect function, ultimately leading to better treatments.”
author and funding
The other author of this study is Gabriel Fitzgerald of Weill Cornell Medicine.Daniel Terry and Zeliha Kirich St. Jude.
This research was supported by the National Institutes of Health (GM079238), Howard Hughes Medical Institute, ALSAC, and funding and advocacy organizations. St. Jude.