title: Structural and functional analysis of mutations in alkaptonuria.
abstract: Alkaptonuria (AKU), the prototypic inborn error of metabolism, was the first human disease to be interpreted as a Mendelian trait by Garrod and Bateson at the beginning of last century. AKU results from impaired function of homogentisate dioxygenase (HGO), an enzyme required for the catabolism of phenylalanine and tyrosine. With the novel 7 AKU and 22 fungal mutations reported here, a total of 84 mutations impairing this enzyme have been found in the HGO gene from humans and model organisms. Forty-three of these mutations result in single amino acid substitutions. This mutational information is analysed here in the context of the HGO structure and function using kinetic assays performed using purified AKU mutant enzymes and the crystal structure of human HGO. HGO is a topologically complex structure which assembles as a functional hexamer arranged as a dimer of trimers. We show how the intricate pattern of intra- and inter-subunit interactions and the extensive surfaces required for subunit folding and association of this oligomeric enzyme can be inactivated at multiple levels by single-residue substitutions. This explains, in part, the predominance of missense mutations (67%) in AKU.
authors.affiliation: Centro de Investigaciones Biológicas CSIC, Velázquez 144, Madrid 28006, Spain.
authors.forename: J M
authors.initials: JM
authors.lastname: Rodríguez
authors.forename: D E
authors.initials: DE
authors.lastname: Timm
authors.forename: G P
authors.initials: GP
authors.lastname: Titus
authors.forename: D
authors.initials: D
authors.lastname: Beltrán-Valero De Bernabé
authors.forename: O
authors.initials: O
authors.lastname: Criado
authors.forename: H A
authors.initials: HA
authors.lastname: Mueller
authors.forename: S
authors.initials: S
authors.lastname: Rodríguez De Córdoba
authors.forename: M A
authors.initials: MA
authors.lastname: Peñalva
mesh_terms: D000474:Alkaptonuria / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001233:Aspergillus nidulans / Q000235:genetics / Q000378:metabolism; D020134:Catalytic Domain; D049308:Dioxygenases; D050560:Homogentisate 1,2-Dioxygenase; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D010105:Oxygenases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011487:Protein Conformation; D017510:Protein Folding; D016415:Sequence Alignment; D013329:Structure-Activity Relationship
chemical_list: D010105:Oxygenases; D049308:Dioxygenases; D050560:Homogentisate 1,2-Dioxygenase