8BCT

X-ray crystal structure of a de novo selected helix-loop-helix heterodimer in a syn arrangement, 26alpha/26beta

Summary for 8BCT

• Entry DOI: 10.2210/pdb8bct/pdb
• Descriptor: 26alpha, 26beta, SULFATE ION, ... (6 entities in total)
• Functional Keywords: coiled coil, 4-helix bundle, in-cell library screening, protein-protein interactions, de novo protein
• Biological source: synthetic construct; More
• Total number of polymer chains: 8
• Total formula weight: 43534.53

Authors

Naudin, E.A.,Mylemans, B.,Smith, A.J.,Savery, N.J.,Woolfson, D.N. (deposition date: 2022-10-17, release date: 2023-06-07, Last modification date: 2024-10-09)

Primary citation

Smith, A.J.,Naudin, E.A.,Edgell, C.L.,Baker, E.G.,Mylemans, B.,FitzPatrick, L.,Herman, A.,Rice, H.M.,Andrews, D.M.,Tigue, N.,Woolfson, D.N.,Savery, N.J.
Design and Selection of Heterodimerizing Helical Hairpins for Synthetic Biology.
Acs Synth Biol, 12:1845-1858, 2023

PubMed 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, 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 and function in cells is not guaranteed. Here, we describe the design, characterization, and application of 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 . 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 and mammalian cells.

PubMed: 37224449
DOI: 10.1021/acssynbio.3c00231

Experimental method

X-RAY DIFFRACTION (1.7 Å)