Tags

Type your tag names separated by a space and hit enter

Hereditary tyrosinemia type I-associated mutations in fumarylacetoacetate hydrolase reduce the enzyme stability and increase its aggregation rate.
J Biol Chem 2019; 294(35):13051-13060JB

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 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

Protein Stability and Inherited Disease Laboratory, CIC bioGUNE, Bizkaia Technology Park, 48160 Derio, Bizkaia, Spain.Protein Stability and Inherited Disease Laboratory, CIC bioGUNE, Bizkaia Technology Park, 48160 Derio, Bizkaia, Spain.Protein Stability and Inherited Disease Laboratory, CIC bioGUNE, Bizkaia Technology Park, 48160 Derio, Bizkaia, Spain.Electron Microscopy Platform, CIC bioGUNE, Bizkaia Technology Park, 48160 Derio, Bizkaia, Spain.Exosomes Laboratory, CIC bioGUNE, Bizkaia Technology Park, 48160 Derio, Bizkaia, Spain.Exosomes Laboratory, CIC bioGUNE, Bizkaia Technology Park, 48160 Derio, Bizkaia, Spain. IKERBASQUE, Basque Foundation for Science, Bilbao, 48013 Spain.Protein Stability and Inherited Disease Laboratory, CIC bioGUNE, Bizkaia Technology Park, 48160 Derio, Bizkaia, Spain omillet@cicbiogune.es.

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, vol. 294, no. 35, 2019, pp. 13051-13060.
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;294(35):13051-13060.
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, 294(35), pp. 13051-13060. 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 Aug 30;294(35):13051-13060. 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 - 2019/05/15/received PY - 2019/07/11/revised PY - 2019/7/14/pubmed PY - 2019/7/14/medline 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 SP - 13051 EP - 13060 JF - The Journal of biological chemistry JO - J. Biol. Chem. VL - 294 IS - 35 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 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 -