Molecular basis of inborn errors of ketone body metabolism:mainly tertiary structural changes of protein and abnormalities of splicing

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The study for molecular basis of inborn errors of ketone body metabolism, especially focused on tertiary structural changes by missense mutations and abnormal splicing, was performed. In collaboration with Finland group, we first showed crystal structure of human mitochondrial acetoacetyl-CoA thiolase (T2) homotetramer. and showed a mechanism of pots ssiurnion activation. We identified mutations in T2 deficiency and succinyl-CoA: 3-ketoacid CoA transferase deficiency. We analyzed 7 new missense mutations by transient expression analysis of cDNA and evaluated temperature sensitive activity and stability. Then we discussed the effect of mutant on the tertiary structure. In case of SCOT deficiency, we identified and characterized a unique splicing mutation. In T2 deficiency, we also identified an exonic mutation which activates cryptic splice donor site just its 5-base upstream. We dearly showed that this exonic mutation was responsible for the aberrant splicing by mini gene splicing experiment. We also showed that alu-mediated genomic rearrangement is one of the cause of T2 deficirncy in some patients.