Projects per year
Abstract
De novo designed protein domains are
increasingly being applied in biotechnology, cell biology and
synthetic biology. Therefore, it is imperative that these proteins
are robust to superficial changes, i.e., small changes to their
amino-acid sequences should not cause gross structural
changes. In turn, this allows properties such as stability and
solubility to be tuned without affecting structural attributes like
tertiary fold and quaternary interactions. Reliably designed
proteins with predictable behaviors may then be used as
scaffolds to incorporate function, e.g., through the introduction
of features for small-molecule, metal or macromolecular
binding, and enzyme-like active sites. Generally, achieving this
requires the starting protein fold to be well understood. Herein,
we focus on designing α-helical coiled coils, which are well
studied, widespread and often direct protein-protein interactions
in natural systems. Our initial investigations reveal that a
previously designed parallel, homotetrameric coiled coil, CCTet, is not robust to sequence changes that were anticipated to
maintain its structure. Instead, the alterations switch the
oligomeric state from tetramer to trimer. To improve the
robustness of designed homotetramers, additional sequences
based on CC-Tet were produced and characterized in solution
and by X-ray crystallography. Of these updated sequences, one
is robust to truncation and to changes in surface electrostatics;
we call this CC-Tet*. Variants of the general CC-Tet* design
provide a set of homotetrameric coiled coils with unfolding
temperatures in the range 40 ˚C to >95 ˚C. We anticipate that
these will be of use in applications requiring robust and well
defined tetramerization domains.
Original language | English |
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Number of pages | 6 |
Journal | Biochemistry |
Early online date | 9 Mar 2020 |
DOIs | |
Publication status | E-pub ahead of print - 9 Mar 2020 |
Structured keywords
- BrisSynBio
- Bristol BioDesign Institute
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Dive into the research topics of 'Robust de novo designed homotetrameric coiled coils'. Together they form a unique fingerprint.Projects
- 3 Finished
-
Design and in vivo assembly of switchable protein-protein interactions for transcription regulation
1/12/18 → 30/11/22
Project: Research
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Design and in vivo assembly of switchable protein-protein interactions for transcription regulation
1/12/18 → 30/11/22
Project: Research
-