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Molecular Mechanism of Localization of the Mutant Ornithine Transcarbamylase to Mitochondrial Compartment

突变型鸟氨酸转氨甲酰酶定位于线粒体区室的分子机制

基本信息

批准号：
09670854
负责人：
YOSHINO Makoto
金额：
$ 1.79万
依托单位：
KURUME UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

项目摘要

1) Research on the transport of wild-type and mutant ornithine transcarbamylase (OTC) into mitochondrial compartmentOTC messenger RNA was isolated from cells transfected with either the wild-type or the R40H OTC cDNA. The mRNA was in vitro-translated with S35, and the labeled OTC was added to isolated rat liver mitochondria. After incubation, incorporation of the label into the protein fractions of various molecular sizes separated on a sephadex column were measured. The incorporation of the label was not reproducible in terms of elution volume and radioactivity in the protein fraction, and further refinements of the experimental conditions were necessary.2) Prenatal monitoring of a pregnancy at risk for OTC deficiency with neonatal onsetThe I261T mutation in the OTC gene was identified in a male neonate who died from hyperammonemic crisis. prenatal diagnosis was done in the successive pregnancy, which revealed that the fetus was female, and that it did not carry the mutant allele. Gene tracking in the family revealed that the mutation was freshly occurred in the mother.3) Analysis of prognostic factors in male patients with late-onset OTC deficiencyPrognosis of life has been generally poor in male patients with the late-onset OTC to deficiency. To elucidate factors that are concerned with prognosis of life, retrospective analysis was done in ten male patients with the disease, none of whom carried the R40H mutation and the other, the Y55D mutation in the OTC gene. A total of 32 factors were first compared in the two groups were further subjected to simple regression analysis, which revealed that age at onset, peak plasma ammonia concentration, concentrations in plasma of eight amino acids were determined to be significant predictors.

1)野生型和突变型鸟氨酸转氨甲酰酶（OTC）转运到线粒体区的研究从转染野生型或R40 H OTC cDNA的细胞中分离OTC信使RNA。用S35体外翻译mRNA，并将标记的OTC加入分离的大鼠肝线粒体中。孵育后，测量标记物掺入在葡聚糖凝胶柱上分离的各种分子大小的蛋白质级分中。标记物的掺入在洗脱体积和蛋白质级分中的放射性方面是不可再现的，并且需要进一步改进实验条件。2）具有OTC缺乏风险的妊娠的产前监测伴新生儿发病在死于高氨血症危象的男性新生儿中鉴定了OTC基因中的I261T突变。在随后的怀孕中进行了产前诊断，结果显示胎儿是女性，并且没有携带突变等位基因。3）男性迟发性OTC缺乏症患者的预后因素分析男性迟发性OTC缺乏症患者的生活预后普遍较差。为了阐明与生命预后有关的因素，对10例男性患者进行了回顾性分析，其中没有一例携带OTC基因的R40H突变，另一例携带OTC基因的Y55D突变。对两组共32个因素进行单因素回归分析，发现发病年龄、血氨峰值浓度、血浆8种氨基酸浓度为显著性预测因素。