title: Functional analysis of MCCA and MCCB mutations causing methylcrotonylglycinuria.
abstract: Methylcrotonylglycinuria (MCG; MIM 210200) is an autosomal recessive inherited human disorder caused by the deficiency of 3-methylcrotonyl-CoA carboxylase (MCC, E.C.6.4.1.4), involved in leucine catabolism. This mitochondrial enzyme is one of the four biotin-dependent carboxylases known in humans. MCC is composed of two different types of subunits, alpha and beta, encoded by the nuclear genes MCCA and MCCB, respectively, recently cloned and characterized. Several mutations have been identified, in both genes, the majority are missense mutations along with splicing mutations and small insertions/deletions. We have expressed four missense mutations, two MCCA and two MCCB mapping to highly evolutionarily conserved residues, by transient transfection of SV40-transformed deficient fibroblasts in order to confirm their pathogenic effect. All the missense mutations expressed resulted in null or severely diminished MCC activity providing direct evidence that they are disease-causing ones. The MCCA mutations have been analysed in the context of three-dimensional structural information modelling the changes in the crystallized biotin carboxylase subunit of the Escherichia coli acetyl-CoA carboxylase. The apparent severity of all the MCC mutations contrasts with the variety of the clinical phenotypes suggesting that there are other cellular and metabolic unknown factors that affect the resulting phenotype.
authors.affiliation: Dpto. Biología Molecular, Centro de Biología Molecular Severo Ochoa CSIC-UAM, Universidad Autónoma de Madrid, Campus de Cantoblanco, Madrid 28049, Spain.
authors.forename: L R
authors.initials: LR
authors.lastname: Desviat
authors.forename: C
authors.initials: C
authors.lastname: Pérez-Cerdá
authors.forename: B
authors.initials: B
authors.lastname: Pérez
authors.forename: J
authors.initials: J
authors.lastname: Esparza-Gordillo
authors.forename: P
authors.initials: P
authors.lastname: Rodríguez-Pombo
authors.forename: M A
authors.initials: MA
authors.lastname: Peñalva
authors.forename: S
authors.initials: S
authors.lastname: Rodríguez De Córdoba
authors.forename: M
authors.initials: M
authors.lastname: Ugarte
mesh_terms: D000595:Amino Acid Sequence; D019735:Carbon-Carbon Ligases / Q000172:deficiency* / Q000235:genetics*; D017124:Conserved Sequence / Q000235:genetics; D005347:Fibroblasts; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D020125:Mutation, Missense / Q000235:genetics*; D020836:Protein Structure, Quaternary; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013539:Simian virus 40; D014162:Transfection
chemical_list: D019735:Carbon-Carbon Ligases; C021573:methylcrotonoyl-CoA carboxylase