title: Hyperinsulinism and hyperammonemia in infants with regulatory mutations of the glutamate dehydrogenase gene.
abstract: BACKGROUND
  A new form of congenital hyperinsulinism characterized by hypoglycemia and hyperammonemia was described recently. We hypothesized that this syndrome of hyperinsulinism and hyperammonemia was caused by excessive activity of glutamate dehydrogenase, which oxidizes glutamate to alpha-ketoglutarate and which is a potential regulator of insulin secretion in pancreatic beta cells and of ureagenesis in the liver.
  
  
  METHODS
  We measured glutamate dehydrogenase activity in lymphoblasts from eight unrelated children with the hyperinsulinism-hyperammonemia syndrome: six with sporadic cases and two with familial cases. We identified mutations in the glutamate dehydrogenase gene by sequencing glutamate dehydrogenase complementary DNA prepared from lymphoblast messenger RNA. Site-directed mutagenesis was used to express the mutations in COS-7 cells.
  
  
  RESULTS
  The sensitivity of glutamate dehydrogenase to inhibition by guanosine 5'-triphosphate was a quarter of the normal level in the patients with sporadic hyperinsulinism-hyperammonemia syndrome and half the normal level in patients with familial cases and their affected relatives, findings consistent with overactivity of the enzyme. These differences in enzyme insensitivity correlated with differences in the severity of hypoglycemia in the two groups. All eight children were heterozygous for the wild-type allele and had a mutation in the proposed allosteric domain of the enzyme. Four different mutations were identified in the six patients with sporadic cases; the two patients with familial cases shared a fifth mutation. In two clones of COS-7 cells transfected with the mutant sequence from one patient, the sensitivity of the enzyme to guanosine 5'-triphosphate was reduced, findings similar to those in the child's lymphoblasts.
  
  
  CONCLUSIONS
  The hyperinsulinism-hyperammonemia syndrome is caused by mutations in the glutamate dehydrogenase gene that impair the control of enzyme activity.
authors.affiliation: Division of Endocrinology, Children's Hospital of Philadelphia, University of Pennsylvania School of Medicine, 19104, USA.
authors.forename: C A
authors.initials: CA
authors.lastname: Stanley
authors.forename: Y K
authors.initials: YK
authors.lastname: Lieu
authors.forename: B Y
authors.initials: BY
authors.lastname: Hsu
authors.forename: A B
authors.initials: AB
authors.lastname: Burlina
authors.forename: C R
authors.initials: CR
authors.lastname: Greenberg
authors.forename: N J
authors.initials: NJ
authors.lastname: Hopwood
authors.forename: K
authors.initials: K
authors.lastname: Perlman
authors.forename: B H
authors.initials: BH
authors.lastname: Rich
authors.forename: E
authors.initials: E
authors.lastname: Zammarchi
authors.forename: M
authors.initials: M
authors.lastname: Poncz
mesh_terms: D000641:Ammonia / Q000097:blood* / Q000378:metabolism; D002648:Child; D002675:Child, Preschool; D004252:DNA Mutational Analysis; D005260:Female; D005969:Glutamate Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D006946:Hyperinsulinism / Q000151:congenital* / Q000201:enzymology / Q000235:genetics; D007223:Infant; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D008297:Male; D008661:Metabolism, Inborn Errors / Q000235:genetics*; D008928:Mitochondria / Q000201:enzymology; D017354:Point Mutation; D013577:Syndrome; D014508:Urea / Q000378:metabolism
chemical_list: D007328:Insulin; D000641:Ammonia; D014508:Urea; D005969:Glutamate Dehydrogenase