Identification of pharmacological chaperones for Phenylalanine Hydroxylase. A virtual screening approach to discover novel drug candidates for treatment of phenylketonuria
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Phenylalanine hydroxylase (PAH) is an enzyme that catalyses the hydroxylation of phenylalanine into tyrosine in the liver, which is the rate-limiting step in phenylalanine catabolism. PAH is a non-heme iron-dependent enzyme that also requires tetrahydrobiopterin (BH4) as cofactor to perform catalysis. PAH dysfunction results in phenylketonuria (PKU), characterized by neurotoxic accumulation of phenylalanine. PKU is caused by mutations in the PAH gene, usually resulting in a misfolding of PAH. A novel approach to treating misfolding diseases in general, is the use of pharmacological chaperones – small molecules that can stabilize the native form of a protein and thus prevent misfolding and rescue protein function. The aim of this Master project was to combine virtual and experimental methods to identify compounds that could act as pharmacological chaperones for PAH. The docking program Glide was used to screen a virtual library, and the compounds with the highest binding affinities were tested experimentally to validate their interactions with PAH, by testing thermostability and enzymatic activity of PAH in the presence of the compounds. Target-based virtual screening and subsequent experimental validation identified a compound with a potential in pharmacological chaperone therapy for PKU. This compound had a significant protective effect on PAH activity, and its specificity and thermodynamic binding properties should be further investigated.