Extension of a de novo TIM barrel with a rationally designed secondary structure element

Titelangaben

Wiese, Jonas Gregor ; Shanmugaratnam, Sooruban ; Höcker, Birte:
Extension of a de novo TIM barrel with a rationally designed secondary structure element.
In: Protein Science. Bd. 30 (Mai 2021) Heft 5 . - S. 982-989.
ISSN 1469-896X
DOI der Verlagsversion: https://doi.org/10.1002/pro.4064

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Abstract

The ability to construct novel enzymes is a major aim in de novo protein design. A popular enzyme fold for design attempts is the TIM barrel. This fold is a common topology for enzymes and can harbor many diverse reactions. The recent de novo design of a four-fold symmetric TIM barrel provides a well understood minimal scaffold for potential enzyme designs. Here we explore opportunities to extend and diversify this scaffold by adding a short de novo helix on top of the barrel. Due to the size of the protein, we developed a design pipeline based on computational ab initio folding that solves a less complex sub-problem focused around the helix and its vicinity and adapt it to the entire protein. We provide biochemical characterization and a high-resolution X-ray structure for one variant and compare it to our design model. The successful extension of this robust TIM-barrel scaffold opens opportunities to diversify it towards more pocket like arrangements and as such can be considered a building block for future design of binding or catalytic sites.