Heparan sulfate proteoglycans play essential roles in forming the extracellular matrix, facilitating protein:protein interactions and acting as a reservoir for signalling molecules. Heparanase is the only mammalian endo-β-glucuronidase is known to catalyse the degradation of heparan sulfate chains and its activity has been linked with a variety of diseases, such as diabetes and cancer. Because of its important role in certain diseases, heparanase is of considerable interest as a drug target. However Despite its biological importance, there has been no successful prokaryotic expression and commercially available heparanase is expressed in mammalian cells is expensive. Insect cell expression has been shown to be possible, but has low stability, poor purity and low yields.
Here we show the use of protein repair one stop shop (PROSS) to design a stable protein in one step. We were able to express heparanase in E.coli and purify to high purity, while retaining catalytic activity, inhibition response and atomic structure.This stabilised version of human heparanase could be used for initial drug screen processes and extends the methods to study which were previously non practical due to limited stability and availability.