Journal
NATURE CHEMISTRY
Volume 8, Issue 9, Pages 837-844Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/NCHEM.2555
Keywords
-
Categories
Funding
- Engineering and Physical Sciences Research Council [EP/G036764/1]
- University of Bristol
- European Research Council [340764]
- Royal Society Wolfson Research Merit Award
- Diamond Light Source [MX-8922]
- Biotechnology and Biological Sciences Research Council [BB/L01386X/1] Funding Source: researchfish
- Engineering and Physical Sciences Research Council [1115150, EP/K03927X/1] Funding Source: researchfish
- BBSRC [BB/L01386X/1] Funding Source: UKRI
- EPSRC [EP/K03927X/1] Funding Source: UKRI
Ask authors/readers for more resources
The design of enzyme-like catalysts tests our understanding of sequence-to-structure/function relationships in proteins. Here we install hydrolytic activity predictably into a completely de novo and thermostable a-helical barrel, which comprises seven helices arranged around an accessible channel. We show that the lumen of the barrel accepts 21 mutations to functional polar residues. The resulting variant, which has cysteine-histidine-glutamic acid triads on each helix, hydrolyses p-nitrophenyl acetate with catalytic efficiencies that match the most-efficient redesigned hydrolases based on natural protein scaffolds. This is the first report of a functional catalytic triad engineered into a de novo protein framework. The flexibility of our system also allows the facile incorporation of unnatural side chains to improve activity and probe the catalytic mechanism. Such a predictable and robust construction of truly de novo biocatalysts holds promise for applications in chemical and biochemical synthesis.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available