Molecular basis of inborn errors of ketone body metabolism, especially succinyl-CoA : 3-ketoacid CoA transferase deficiency

1)SCOT缺陷：我们揭示了人SCOT基因的结构和序列，建立了人SCOT蛋白的三级结构模型，并鉴定和表征了3例SCOT缺陷患者的基因突变。我们现在可以在基因组水平上筛选SCOT基因突变。我们分析了人SCOT基因的5'侧翼区，以阐明肝细胞特异性抑制SCOT基因的机制。我们测定了该区域3 kb的序列，发现Sp1基本上驱动SCOT基因的表达。2）T2缺乏：我们在T2蛋白三级结构模型上分析了5例西班牙T2缺乏患者的氨基酸改变（基因突变）特征。在一例患者中，我们发现了一个新的剪接突变380 C>T，该突变位于外显子3上，激活了外显子3上预先存在的隐蔽剪接位点，导致隐蔽位点的排他性剪接。我们在26例T2缺陷患者中发现了基因突变。我们收集了这些患者的临床信息，使用问卷调查，以搜索临床过程和结果，并澄清T2缺乏症的基因型/表型相关性。通过本研究，我们得出以下结论：（1）T2缺乏总体上具有良好的结局;（2）在确诊后可以避免严重的酮症酸中毒发作;（3）没有明显的表型/基因型相关性。首席研究员荣幸地在“酶学方法”和“遗传性疾病的代谢和分子基础”中撰写章节。

1) SCOT deficieicny : We revealed the structure and sequence of human SCOT gene, made tertiary structural model of human SCOT protein, and identified and characterized gene mutations in 3 SCOT deficient patients. We now can screen SCOT gene mutations at the genomic level. We analyzed 5' flanking regions of human SCOT gene in order to clarify the mechanism for specific SCOT gene supression in hepatocytes. We determined the sequence of 3 kb in the region, and revealed that Sp1 basically drive the SCOT gene expression. We also searched cis-elements responsible for specific supression in hepatocytes.2) T2 deficiency : We analyzed character of amino acid alternations (gene mutations) identified in 5 spanish T2 deficient patients on teritiary structural model of T2 protein. In one patient, we identified a novel splicing mutation 380C>T.This mutation located on exon 3 activated pre-exsisting cryptic splice site on exon 3, resulting in exclusive splicing at the cryptic site. We have identified gene mutations in 26 T2 deficient patients. We collected clinical information from the physicians for these patients with the use of questionnaire to search clinical courses and outcome and to clarifiy genotype/phenotype correlation in T2 deficiency. By the study, we concluded that (1) T2 deficiency has a favorable outcome in general ; (2) severe ketoacidotic attack can be avoidable after confirmation of the diagnosis ; (3) there is no apparent phenotype/genotype correlation.The head investigator was honored to write chapters in "Methods in Enzymology" and in "Metabolic ＆ Molecular Bases of Inherited Disease".