Issue 3, 2022

Computational design and experimental characterisation of a stable human heparanase variant

Abstract

Heparanase is the only human enzyme known to hydrolyse heparin sulfate and is involved in many important physiological processes. However, it is also unregulated in many disease states, such as cancer, diabetes and Covid-19. It is thus an important drug target, yet the heterologous production of heparanase is challenging and only possible in mammalian or insect expression systems, which limits the ability of many laboratories to study it. Here we describe the computational redesign of heparanase to allow high yield expression in Escherchia coli. This mutated form of heparanase exhibits essentially identical kinetics, inhibition, structure and protein dynamics to the wild type protein, despite the presence of 26 mutations. This variant will facilitate wider study of this important enzyme and contributes to a growing body of literature that shows evolutionarily conserved and functionally neutral mutations can have significant effects on protein folding and expression.

Graphical abstract: Computational design and experimental characterisation of a stable human heparanase variant

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Article information

Article type
Paper
Submitted
13 12月 2021
Accepted
11 2月 2022
First published
15 2月 2022
This article is Open Access
Creative Commons BY-NC license

RSC Chem. Biol., 2022,3, 341-349

Computational design and experimental characterisation of a stable human heparanase variant

C. Whitefield, N. Hong, J. A. Mitchell and C. J. Jackson, RSC Chem. Biol., 2022, 3, 341 DOI: 10.1039/D1CB00239B

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