title: Isolated 2-methylbutyrylglycinuria caused by short/branched-chain acyl-CoA dehydrogenase deficiency: identification of a new enzyme defect, resolution of its molecular basis, and evidence for distinct acyl-CoA dehydrogenases in isoleucine and valine metabolism.
abstract: Acyl-CoA dehydrogenase (ACAD) defects in isoleucine and valine catabolism have been proposed in clinically diverse patients with an abnormal pattern of metabolites in their urine, but they have not been proved enzymatically or genetically, and it is unknown whether one or two ACADs are involved. We investigated a patient with isolated 2-methylbutyrylglycinuria, suggestive of a defect in isoleucine catabolism. Enzyme assay of the patient's fibroblasts, using 2-methylbutyryl-CoA as substrate, confirmed the defect. Sequence analysis of candidate ACADs revealed heterozygosity for the common short-chain ACAD A625 variant allele and no mutations in ACAD-8 but a 100-bp deletion in short/branched-chain ACAD (SBCAD) cDNA from the patient. Our identification of the SBCAD gene structure (11 exons; >20 kb) enabled analysis of genomic DNA. This showed that the deletion was caused by skipping of exon 10, because of homozygosity for a 1228G-->A mutation in the patient. This mutation was not present in 118 control chromosomes. In vitro transcription/translation experiments and overexpression in COS cells confirmed the disease-causing nature of the mutant SBCAD protein and showed that ACAD-8 is an isobutyryl-CoA dehydrogenase and that both wild-type proteins are imported into mitochondria and form tetramers. In conclusion, we report the first mutation in the SBCAD gene, show that it results in an isolated defect in isoleucine catabolism, and indicate that ACAD-8 is a mitochondrial enzyme that functions in valine catabolism.
authors.affiliation: Research Unit for Molecular Medicine, Aarhus University Hospital, and Faculty of Health Science, Skejby Sygehus, DK 8200 Arhus N, Denmark. brage@biobase.dk
authors.forename: B S
authors.initials: BS
authors.lastname: Andresen
authors.forename: E
authors.initials: E
authors.lastname: Christensen
authors.forename: T J
authors.initials: TJ
authors.lastname: Corydon
authors.forename: P
authors.initials: P
authors.lastname: Bross
authors.forename: B
authors.initials: B
authors.lastname: Pilgaard
authors.forename: R J
authors.initials: RJ
authors.lastname: Wanders
authors.forename: J P
authors.initials: JP
authors.lastname: Ruiter
authors.forename: H
authors.initials: H
authors.lastname: Simonsen
authors.forename: V
authors.initials: V
authors.lastname: Winter
authors.forename: I
authors.initials: I
authors.lastname: Knudsen
authors.forename: L D
authors.initials: LD
authors.lastname: Schroeder
authors.forename: N
authors.initials: N
authors.lastname: Gregersen
authors.forename: F
authors.initials: F
authors.lastname: Skovby
mesh_terms: D017398:Alternative Splicing / Q000235:genetics; D000592:Amino Acid Metabolism, Inborn Errors / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002675:Child, Preschool; D003241:Consanguinity; D004252:DNA Mutational Analysis; D004795:Enzyme Stability; D005091:Exons; D005260:Female; D006579:Heterozygote; D006801:Humans; D007438:Introns; D007532:Isoleucine / Q000378:metabolism*; D008297:Male; D008928:Mitochondria / Q000201:enzymology / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D010088:Oxidoreductases / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D044925:Oxidoreductases Acting on CH-CH Group Donors; D010154:Pakistan; D021381:Protein Transport; D017384:Sequence Deletion / Q000235:genetics; D014162:Transfection; D014633:Valine / Q000378:metabolism*
chemical_list: D007532:Isoleucine; D010088:Oxidoreductases; D044925:Oxidoreductases Acting on CH-CH Group Donors; C038808:2-methylacyl-CoA dehydrogenase; D014633:Valine