Biochemical and cellular studies of three human 3‐phosphoglycerate dehydrogenase variants responsible for pathological reduced L‐serine levels

Abstract

AbstractIn the brain, the non‐essential amino acid L‐serine is produced through the phosphorylated pathway (PP) starting from the glycolytic intermediate 3‐phosphoglycerate: among the different roles played by this amino acid, it can be converted into D‐serine and glycine, the two main co‐agonists of NMDA receptors. In humans, the enzymes of the PP, namely phosphoglycerate dehydrogenase (hPHGDH, which catalyzes the first and rate‐limiting step of this pathway), 3‐phosphoserine aminotransferase, and 3‐phosphoserine phosphatase are likely organized in the cytosol as a metabolic assembly (a “serinosome”). The hPHGDH deficiency is a pathological condition biochemically characterized by reduced levels of L‐serine in plasma and cerebrospinal fluid and clinically identified by severe neurological impairment. Here, three single‐point variants responsible for hPHGDH deficiency and Neu‐Laxova syndrome have been studied. Their biochemical characterization shows that V261M, V425M, and V490M substitutions alter either the kinetic (both maximal activity and Km for 3‐phosphoglycerate in the physiological direction) and the structural properties (secondary, tertiary, and quaternary structure, favoring aggregation) of hPHGDH. All the three variants have been successfully ectopically expressed in U251 cells, thus the pathological effect is not due to hindered expression level. At the cellular level, mistargeting and aggregation phenomena have been observed in cells transiently expressing the pathological protein variants, as well as a reduced L‐serine cellular level. Previous studies demonstrated that the pharmacological supplementation of L‐serine in hPHGDH deficiencies could ameliorate some of the related symptoms: our results now suggest the use of additional and alternative therapeutic approaches.