Insights into the pathogenesis of primary hyperoxaluria type I from the structural dynamics of alanine:glyoxylate aminotransferase variants

Primary hyperoxaluria type I (PH1) is caused by deficient alanine:glyoxylate aminotransferase (AGT) activity. PH1-causing mutations in AGT lead to protein mistargeting and aggregation. Here, we use hydrogen-deuterium exchange (HDX) to characterize the wild-type (WT), the LM (a polymorphism frequent in PH1 patients) and the LM G170R (the most common mutation in PH1) variants of AGT. We provide the first experimental analysis of AGT structural dynamics, showing that stability is heterogeneous in the native state and providing a blueprint for frustrated regions with potentially functional relevance. The LM and LM G170R variants only show local destabilization. Enzymatic transamination of the pyridoxal 5-phosphate cofactor bound to AGT hardly affects stability. Our study, thus, supports that AGT misfolding is not caused by dramatic effects on structural dynamics. Mutations in alanine:glyoxylate aminotransferase (AGT) leading to protein misfolding cause primary hyperoxaluria type I (PH1). Here, we use hydrogen-deuterium exchange to characterize the structural dynamics of AGT variants. These dynamics are heterogeneously distributed across the structure and different functional sites. Disease-associated variants show local destabilization. Our work provides novel insight into the pathogenic mechanisms causing PH1.image