Molecular characterization of 82 patients with pyruvate dehydrogenase complex deficiency. Structural implications of novel amino acid substitutions in E1 protein
Molecular characterization of 82 patients with pyruvate dehydrogenase complex deficiency. Structural implications of novel amino acid substitutions in E1 protein
复制标题
DOI：
10.1016/j.ymgme.2011.08.008
复制
发表时间：
2011-12-01
影响因子：
3.8
通讯作者：
Brivet, M.
中科院分区：
文献类型：
--
作者：
Imbard, A.;Boutron, A.;Brivet, M.
背景：丙酮酸脱氢酶复合物（PDHc）缺乏是原发性乳酸酸中毒的重要原因。大多数病例是由X连锁的丙酮酸脱氢酶E1 α亚基（PDHA 1）基因突变引起的，而少数病例是由E1 β（PDHB）、E2（DLAT）、E3（DLD）和E3 BP（PDHX）亚基或PDH磷酸酶（PDP 1）基因突变引起的。目的：报道82例PDH c缺陷患者的分子特征，分析PDHA 1新错义突变的结构效应。首先，通过使用长范围PCR产物的外显子测序、基因剂量测定和cDNA分析来研究PDHA 1变异。在未解决的病例中进一步进行PDHX、PDHB、DLAT和DLD cDNA的突变扫描。结果：在30例女孩和35例男孩中发现PDHA 1基因突变。三个大的重排，包括两个连续的基因缺失综合征被确定。新的错义，移码和剪接突变也划定和无义突变的镶嵌男性。PDHA 1中的突变p.G1u75Ala、p.Arg88Ser、p.Arg119Trp、p.Gly144Asp、p.Pro217Arg、p.Arg235Gly、p.Tyr243Cys、p.Tyr243Ser、p.Arg245Gly、p.Pro250Leu、p.Gly278Arg、p.Met282Val、p.Gly298Glu被预测为损害活性位点通道构象或亚基相互作用。7例PDHB突变患者中有6例显示复发性p.Met101Val突变; 9例患者携带PDHX突变和1例患者DLD突变。结论：我们提供了一种有效的逐步策略，用于PDHc基因突变筛查，并扩展了在结构水平上分析的PDHA 1突变的不断增长的列表。(C)2011 Elsevier Inc. All rights reserved.
Background: Pyruvate dehydrogenase complex (PDHc) deficiencies are an important cause of primary lactic acidosis. Most cases result from mutations in the X-linked gene for the pyruvate dehydrogenase E1 alpha subunit (PDHA1) while a few cases result from mutations in genes for E1 beta (PDHB), E2 (DLAT), E3 (DLD) and E3BP (PDHX) subunits or PDH-phosphatase (PDP1).Aim: To report molecular characterization of 82 PDHc-deficient patients and analyze structural effects of novel missense mutations in PDHA1.Methods: PDHA1 variations were investigated first, by exon sequencing using a long range PCR product, gene dosage assay and cDNA analysis. Mutation scanning in PDHX, PDHB, DLAT and DLD cDNAs was further performed in unsolved cases. Novel missense mutations in PDHA1 were located on the tridimensional model of human E1 protein to predict their possible functional consequences.Results: PDHA1 mutations were found in 30 girls and 35 boys. Three large rearrangements, including two contiguous gene deletion syndrome were identified. Novel missense, frameshift and splicing mutations were also delineated and a nonsense mutation in a mosaic male. Mutations p.G1u75Ala, p.Arg88Ser, p.Arg119Trp, p.Gly144Asp, p.Pro217Arg, p.Arg235Gly, p.Tyr243Cys, p.Tyr243Ser, p.Arg245Gly, p.Pro250Leu, p.Gly278Arg, p.Met282Val, p.Gly298Glu in PDHA1 were predicted to impair active site channel conformation or subunit interactions. Six out of the seven patients with PDHB mutations displayed the recurrent p.Met101Val mutation; 9 patients harbored PDHX mutations and one patient DLD mutations.Conclusion: We provide an efficient stepwise strategy for mutation screening in PDHc genes and expand the growing list of PDHA1 mutations analyzed at the structural level. (C) 2011 Elsevier Inc. All rights reserved.