Abstract

This talk will describe how we have developed an understanding of natural protein-folding motif, namely the alpha-helical coiled-coil that directs protein oligomerization; and then how we can apply this knowledge in rational peptide and protein design. For the latter, it is now clear that nature has almost certainly only shown us a glimpse of what is possible structurally and functionally with polypeptide chains. To help us explore past the confines of natural coiled-coil structures, and into what has been termed the dark matter of protein space, we have developed a toolkit of de novo coiled-coil peptides (1). These can be used as building blocks for the rapid construction of new protein structures and assemblies. The talk will demonstrate the utility of this approach to make water-soluble protein-like barrels and pores (2, 3), which can be engineered to bind biological molecules and catalyse simple reactions (4, 5). More recently we have engineered membrane-spanning variants of these alpha-helical barrels, and in collaboration with Hagan Bayley’s lab, we have shown that these insert into lipid bilayers and conduct ions in voltage-dependent manners. The insertion and orientation of peptides into the membranes can be directed by design, and the channel activities can be controlled through mutagenesis and blocking reagents.

1. A Basis Set of de Novo Coiled-Coil Peptide Oligomers for Rational Protein Design and Synthetic Biology JM Fletcher, AL Boyle, M Bruning, GJ Bartlett, TL Vincent, NR Zaccai, CT Armstrong, EHC Bromley, PJ Booth, RL Brady, AR Thomson, and DN Woolfson ACS Synthetic Biology 6, 240-250 (2012)

2. A de novo peptide hexamer with a mutable channel NR Zaccai, B Chi, AR Thomson, AL Boyle, GJ Bartlett, M Bruning, N Linden, RB Sessions, PJ Booth, RL Brady and DN Woolfson Nature Chemical Biology 7, 935-941 (2011)

3. Computational design of water-soluble alpha-helical barrels AR Thomson, CW Wood, AJ Burton, GJ Bartlett, RB Sessions, RL Brady and DN Woolfson Science 346, 485-488 (2014)

4. Accessibility, Reactivity, and Selectivity of Side Chains within a Channel of de Novo Peptide Assembly AJ Burton, F Thomas, C Agnew, KL Hudson, SE Halford, RL Brady and DN Woolfson J Am Chem Soc 135, 12524-12527 (2013).

5. Modular Design of Self-Assembling Peptide-Based Nanotubes. NC Burgess, TH Sharp, F Thomas, CW Wood, AR Thomson, NR Zaccai, RL Brady, LC Serpell and DN Woolfson J Am Chem Soc 137, 10554-10562 (2015).