Functional Mapping and Engineering of the Sec Translocon Unlocked by a Cell-Free System
Meier, M.; Scholz, S. A.; von Bank, L.; Levandoski, J. E.; Lückhof, M.; Lieberwirth, I.; Landfester, K.; Jung, A. L.; Schaaf, M.; Si, S.; Driessen, A. J. M.; Erb, T. J.
Show abstract
Almost all proteins are inserted or translocated across membranes by the universally conserved Sec translocon. Despite its central role, experimental access to Sec function has remained limited. Here, we present a cell-free protein synthesis platform that inserts SecYEG into synthetic vesicles, enabling direct testing of Sec in real-time and high-throughput, circumventing longstanding viability constraints. Screening 300 Sec variants in a single experiment, we consolidate three decades of Sec research, while vastly expanding mutant diversity for structure-function insights. Mapping over 30 functionally critical regions that modulate Sec activity across three orders of magnitude, we uncover dozens of super-active variants. We further leverage our system to increase membrane protein quality and nanobody export, highlighting the potential of our system for advancing applications in synthetic biology and biotechnology.
Matching journals
The top 3 journals account for 50% of the predicted probability mass.
Similar papers in this journal
Similar papers in this journal
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.