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Instruments / Technical Area	Publication Details	Published	Actions
I04-Macromolecular Crystallography	Design and selection of heterodimerizing helical hairpins for synthetic biology Abigail J. Smith , Elise A. Naudin , Caitlin L. Edgell , Emily G. Baker , Bram Mylemans , Laura Fitzpatrick , Andrew Herman , Helen M. Rice , David M. Andrews , Natalie Tigue , Derek N. Woolfson , Nigel J. Savery Acs Synthetic Biology DOI: 10.1021/acssynbio.3c00231 Diamond Proposal Number(s): [23269] Open Access Show Abstract ▼ Abstract: Synthetic biology applications would benefit from protein modules of reduced complexity that function orthogonally to cellular components. As many subcellular processes depend on peptide–protein or protein–protein interactions, de novo designed polypeptides that can bring together other proteins controllably are particularly useful. Thanks to established sequence-to-structure relationships, helical bundles provide good starting points for such designs. Typically, however, such designs are tested in vitro and function in cells is not guaranteed. Here, we describe the design, characterization, and application of de novo helical hairpins that heterodimerize to form 4-helix bundles in cells. Starting from a rationally designed homodimer, we construct a library of helical hairpins and identify complementary pairs using bimolecular fluorescence complementation in E. coli. We characterize some of the pairs using biophysics and X-ray crystallography to confirm heterodimeric 4-helix bundles. Finally, we demonstrate the function of an exemplar pair in regulating transcription in both E. coli and mammalian cells.	May 2023

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I04-Macromolecular Crystallography

Design and selection of heterodimerizing helical hairpins for synthetic biology