title: Molecular study of electron transfer flavoprotein alpha-subunit deficiency in two Japanese children with different phenotypes of glutaric acidemia type II.
abstract: BACKGROUND
  Electron transfer flavoprotein is a mitochondrial matrix protein composed of alpha- and beta-subunits (ETF alpha and ETF beta, respectively). This protein transfers electrons between several mitochondrial dehydrogenases and the main respiratory chain via ETF dehydrogenase (ETF-DH). Defects in ETF or ETF-DH cause glutaric acidemias type II (GAII).
  
  
  METHODS
  We investigated the molecular basis of ETF alpha deficiency in two Japanese children with different clinical phenotypes using expression study.
  
  
  RESULTS
  Patient 1 had the severe form of GAII, a compound heterozygote of two mutations: 799G to A (alpha G267R) and nonsense 7C to T (alpha R3X). Patient 2 had the mild form and carried two heterozygous mutations: 764G to T (alpha G255V) and 478delG (frameshift). Both patients had one each of missense mutations in one allele; the others were either nonsense or truncated. Restriction enzyme digestion assay using genomic DNAs from 100 healthy Japanese revealed that these mutations were all novel. No signal for ETF alpha was detected by immunoblotting in cases of missense mutants, while wild-type cDNA resulted in expression of ETF alpha protein. Transfection with wild-type ETF alpha cDNA into cultured cells from both patients elevated incorporation of radioisotope-labelled fatty acids.
  
  
  CONCLUSIONS
  These four mutations were pathogenic for GAII and missense mutations, alpha G255V and alpha G267R were considered anecdotal for mild and severe forms, respectively.
authors.affiliation: Department of Pediatric, Shimane Medical University, Izumo, Shimane, Japan.
authors.forename: E
authors.initials: E
authors.lastname: Purevjav
authors.forename: M
authors.initials: M
authors.lastname: Kimura
authors.forename: Y
authors.initials: Y
authors.lastname: Takusa
authors.forename: T
authors.initials: T
authors.lastname: Ohura
authors.forename: M
authors.initials: M
authors.lastname: Tsuchiya
authors.forename: N
authors.initials: N
authors.lastname: Hara
authors.forename: T
authors.initials: T
authors.lastname: Fukao
authors.forename: S
authors.initials: S
authors.lastname: Yamaguchi
mesh_terms: D044927:Electron-Transferring Flavoproteins; D005227:Fatty Acids / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D005420:Flavoproteins / Q000235:genetics* / Q000378:metabolism*; D016368:Frameshift Mutation; D005977:Glutarates / Q000097:blood*; D006801:Humans; D007231:Infant, Newborn; D007564:Japan; D008297:Male; D008661:Metabolism, Inborn Errors / Q000175:diagnosis / Q000235:genetics*; D009154:Mutation; D020125:Mutation, Missense; D010641:Phenotype; D017354:Point Mutation; D020033:Protein Isoforms / Q000172:deficiency / Q000235:genetics
chemical_list: D044927:Electron-Transferring Flavoproteins; D005227:Fatty Acids; D005420:Flavoproteins; D005977:Glutarates; D020033:Protein Isoforms