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Hereditary tyrosinemia type I-associated mutations in fumarylacetoacetate hydrolase reduce the enzyme stability and increase its aggregation rate.

Abstract

More than 100 mutations in the gene encoding fumarylacetoacetate hydrolase (FAH) cause hereditary tyrosinemia type I (HT1), a metabolic disorder characterized by elevated blood levels of tyrosine. Some of these mutations are known to decrease FAH catalytic activity, but the mechanisms of FAH mutation-induced pathogenicity remain poorly understood. Here, using diffusion ordered NMR spectroscopy, cryo-EM, and CD analyses, along with site-directed mutagenesis, enzymatic assays, and molecular dynamics simulations, we investigated the putative role of thermodynamic and kinetic stability in wild type (WT) FAH and a representative set of 19 missense mutations identified in individuals with HT1. We found that at physiological temperatures and concentrations, WT FAH is in equilibrium between a catalytically active dimer and a monomeric species, with the latter being inactive and prone to oligomerization and aggregation. We also found that the majority of the deleterious mutations reduce the kinetic stability of the enzyme and always accelerate the FAH aggregation pathway. Depending mainly on the position of the amino acid in the structure, pathogenic mutations either reduced the dimer population or decreased the energy barrier that separates the monomer from the aggregate. The mechanistic insights reported here pave the way for the development of pharmacological chaperones that target FAH to tackle the severe disease HT1.

Authors+Show Affiliations

CIC bioGUNE, Spain.CIC bioGUNE.CIC bioGUNE, Spain.CIC bioGUNE.CIC bioGUNE, CIBERehd.CIC bioGUNE, Spain.CIC bioGUNE, Spain.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31300554

Citation

Macias, Iratxe, et al. "Hereditary Tyrosinemia Type I-associated Mutations in Fumarylacetoacetate Hydrolase Reduce the Enzyme Stability and Increase Its Aggregation Rate." The Journal of Biological Chemistry, 2019.
Macias I, Laín A, Bernardo-Seisdedos G, et al. Hereditary tyrosinemia type I-associated mutations in fumarylacetoacetate hydrolase reduce the enzyme stability and increase its aggregation rate. J Biol Chem. 2019.
Macias, I., Laín, A., Bernardo-Seisdedos, G., Gil, D., Gonzalez, E., Falcon-Perez, J. M., & Millet, O. (2019). Hereditary tyrosinemia type I-associated mutations in fumarylacetoacetate hydrolase reduce the enzyme stability and increase its aggregation rate. The Journal of Biological Chemistry, doi:10.1074/jbc.RA119.009367.
Macias I, et al. Hereditary Tyrosinemia Type I-associated Mutations in Fumarylacetoacetate Hydrolase Reduce the Enzyme Stability and Increase Its Aggregation Rate. J Biol Chem. 2019 Jul 12; PubMed PMID: 31300554.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Hereditary tyrosinemia type I-associated mutations in fumarylacetoacetate hydrolase reduce the enzyme stability and increase its aggregation rate. AU - Macias,Iratxe, AU - Laín,Ana, AU - Bernardo-Seisdedos,Ganeko, AU - Gil,David, AU - Gonzalez,Esperanza, AU - Falcon-Perez,Juan M, AU - Millet,Oscar, Y1 - 2019/07/12/ PY - 2019/07/12/accepted PY - 2019/05/15/received PY - 2019/7/14/entrez KW - biophysics KW - enzyme mutation KW - fumaryl acetoacetate hydrolase KW - nuclear magnetic resonance (NMR) KW - protein aggregation KW - protein stability KW - rare disease KW - tyrosine KW - tyrosinemia type I JF - The Journal of biological chemistry JO - J. Biol. Chem. N2 - More than 100 mutations in the gene encoding fumarylacetoacetate hydrolase (FAH) cause hereditary tyrosinemia type I (HT1), a metabolic disorder characterized by elevated blood levels of tyrosine. Some of these mutations are known to decrease FAH catalytic activity, but the mechanisms of FAH mutation-induced pathogenicity remain poorly understood. Here, using diffusion ordered NMR spectroscopy, cryo-EM, and CD analyses, along with site-directed mutagenesis, enzymatic assays, and molecular dynamics simulations, we investigated the putative role of thermodynamic and kinetic stability in wild type (WT) FAH and a representative set of 19 missense mutations identified in individuals with HT1. We found that at physiological temperatures and concentrations, WT FAH is in equilibrium between a catalytically active dimer and a monomeric species, with the latter being inactive and prone to oligomerization and aggregation. We also found that the majority of the deleterious mutations reduce the kinetic stability of the enzyme and always accelerate the FAH aggregation pathway. Depending mainly on the position of the amino acid in the structure, pathogenic mutations either reduced the dimer population or decreased the energy barrier that separates the monomer from the aggregate. The mechanistic insights reported here pave the way for the development of pharmacological chaperones that target FAH to tackle the severe disease HT1. SN - 1083-351X UR - https://www.unboundmedicine.com/medline/citation/31300554/Hereditary_tyrosinemia_type_I-associated_mutations_in_fumarylacetoacetate_hydrolase_reduce_the_enzyme_stability_and_increase_its_aggregation_rate L2 - http://www.jbc.org/cgi/pmidlookup?view=long&pmid=31300554 DB - PRIME DP - Unbound Medicine ER -