title: Anabolic function of phenylalanine hydroxylase in Caenorhabditis elegans.
abstract: In humans, liver phenylalanine hydroxylase (PAH) has an established catabolic function, and mutations in PAH cause phenylketonuria, a genetic disease characterized by neurological damage, if not treated. To obtain novel evolutionary insights and information on molecular mechanisms operating in phenylketonuria, we investigated PAH in the nematode Caenorhabditis elegans (cePAH), where the enzyme is coded by the pah-1 gene, expressed in the hypodermis. CePAH presents similar molecular and kinetic properties to human PAH [S(0.5)(L-Phe) approximately 150 microM; K(m) for tetrahydrobiopterin (BH(4)) approximately 35 microM and comparable V(max)], but cePAH is devoid of positive cooperativity for L-Phe, an important regulatory mechanism of mammalian PAH that protects the nervous system from excess L-Phe. Pah-1 knockout worms show no obvious neurological defects, but in combination with a second cuticle synthesis mutation, they display serious cuticle abnormalities. We found that pah-1 knockouts lack a yellow-orange pigment in the cuticle, identified as melanin by spectroscopic techniques, and which is detected in C. elegans for the first time. Pah-1 mutants show stimulation of superoxide dismutase activity, suggesting that cuticle melanin functions as oxygen radical scavenger. Our results uncover both an important anabolic function of PAH and the change in regulation of the enzyme along evolution.
authors.affiliation: Department of Biomedicine, University of Bergen, Jonas Lies vei 91, 5009 Bergen, Norway.
authors.forename: Ana C
authors.initials: AC
authors.lastname: Calvo
authors.forename: Angel L
authors.initials: AL
authors.lastname: Pey
authors.forename: Ming
authors.initials: M
authors.lastname: Ying
authors.forename: Curtis M
authors.initials: CM
authors.lastname: Loer
authors.forename: Aurora
authors.initials: A
authors.lastname: Martinez
mesh_terms: D045930:Anabolic Agents / Q000378:metabolism; D000818:Animals; D005075:Biological Evolution; D017173:Caenorhabditis elegans / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism; D016166:Free Radical Scavengers / Q000378:metabolism; D017353:Gene Deletion; D017238:Genes, Helminth; D007700:Kinetics; D008543:Melanins / Q000378:metabolism; D009997:Osmotic Pressure; D018384:Oxidative Stress; D010641:Phenotype; D010651:Phenylalanine Hydroxylase / Q000737:chemistry / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation; D013379:Substrate Specificity; D013381:Subtilisins / Q000235:genetics / Q000378:metabolism
chemical_list: D045930:Anabolic Agents; D029742:Caenorhabditis elegans Proteins; D016166:Free Radical Scavengers; D008543:Melanins; D010651:Phenylalanine Hydroxylase; D013381:Subtilisins; C093968:bli-4 protein, C elegans