Participants
Shi Yuan Ng
Classic galactosaemia is a rare metabolic disorder caused by a deficiency in galactose-1-phosphate uridylyltransferase, leading to the toxic accumulation of galactose-1-phosphate. Inhibiting galactokinase 1 (GALK1)—the upstream enzyme that converts galactose into galactose-1-phosphate — could potentially reduce this accumulation. Previous research has found that GALK1 can be inhibited via an allosteric site. However, the precise mode of action is still unclear.
My project aimed to determine the effect of 5 allosteric site mutations on GALK1 structure and function. The kinase activity of the variants was screened with Kinase Glo assay and the protein stability and folding were also measured with NanoDSF. Despite no significant effect on the kinase activity, a reduction in protein stability was oberserved for all variants. All variants crystallised and data was collected at medium resolution. I have solved one of the variants’ structure and we found the overall structure is in agreement with the AlphaFold model.