Milosevic, S.; Cano, M.; Salido, E.; Mesa, N.; Pey, A. l.

Phosphorylation is fundamental to modulate protein function and stabilty. There have detected about 300000 site-specic Phosphorylation sites in over 20000 human proteins. However, only 5% of the sites have been experimenrally characterized. In this work, we investigated a phosphorylation event in AGT, an important enzyme due to its detoxifying role of glyoxylate and hundreds of mutations cause a rare disease (primary hyperoxaluria type I or PH1). We analyzed the effect of phosphomimetic mutations Ser81 on the WT proten, the common polymorpshim LM, and the most common disease-associted variants (LM-G170R and LM-I244T). Using biochemical, biophysical and cell biology approaches, we show that phosphorylation at S81 dramatically affects PLP/PMP binding pose and disrupts enzyme activity, without pertubing its subcellular location to peroxisomes. This reversible phenotype is similar to the irreversible effects of some PH1-causing mutaions. Thus, we provide evidence for a novel regulatory mechanism for PH1, in health and disease.