The Landscape of Mutations in Human Fumarate Hydratase

ABSTRACT Fumarate Hydratase (FH) is an enzyme of the citric acid (TCA) cycle that is responsible for reversibly catalysing the conversion between fumarate and malate. FH loss and subsequent buildup of the oncometabolite fumarate causes hereditary leiomyomatosis and renal cell carcinoma. We explore the mutational landscape of FH in silico, predict the functional effects of many already detected mutations, and categorise detected but un-characterised mutations in human populations. Using mutational energy predicting tools such as Rosetta and FoldX we accurately predict mutations and mutational hotspots with high disruptive capability. Furthermore, through performing molecular dynamics simulations we show that hinge regions of the protein can be stabilized or destabilized by mutations, with new mechanistic implications of the consequences on the binding affinity of the enzyme for its substrates. Finally, we categorise all potential mutations in FH into functional groups, and predict which known mutations in the human population are loss-of-function, and therefore predispose patients to papillary renal carcinoma – we validate our findings through analysis of metabolomics data of characterized cell lines.