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Molecular and cellular analysis of peptidase D (prolidase) deficiency.

肽酶 D（脯氨酸酶）缺乏症的分子和细胞分析。

基本信息

批准号：
02671047
负责人：
ENDO Fumio
金额：
$ 1.41万
依托单位：
Kumamoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02671047/
关键词：
peptidase D prolidase inborn errors of metabolism gene analysis amino acid metabolism autosomal recessive プリダ-ゼ筋ジストロフィ-遺伝子筋緊張型筋ジストロフィ-

项目摘要

Peptidase D (prolidase) deficiency is a rare autosomal recessive disorder characterized by various skin lesions, mental retardation, and other symptom. Peptidase D (EC 3.4.13.9) is a kind of oligopeptidases which hydrolysis imidopeptides and releases carboxyl terminal proline from the substrate peptides. We have isolated the enzyme from human blood and cloned the CDNA for human peptidase D. Then we have determined the primary structure of the enzyme, the structure of the gene and the localization of the gene.We carried out the gene analysis of the patient with the disease. In a Japanese family with two sisters with the-disease, we found 192base pairs (bp) deflection in MRNA from cultured cells. We cloned the gene fragment which covered the deletion mutation from the patients and determined the nucleotide sequences. It was demonstrated that approximately 759 bp sequence was deleted in the gene from the patients. In the case from Belgium (originally from Meddle East), we found a point mutation (A to G at nucleotide 826) which resulted in an amino acid substitution (Asp-Asn). Expression analysis of the mutant CDNA revealed that the inactive synthesized from the CDNA. The same mutation was found in the other patient who came from Middle East. Thus the point mutation might be originated from the Middle East.In addition we determined the nucleotide sequences surrounding the intron-exon junctions. These studies make us possible to amplify the all exons for gene analysis of this disease.

肽酶D（脯氨酸酶）缺乏症是一种罕见的常染色体隐性遗传病，以各种皮肤病变、智力低下等症状为特征。肽酶 D (EC 3.4.13.9) 是一种寡肽酶，可水解酰亚胺肽并从底物肽中释放羧基末端脯氨酸。我们从人的血液中分离出了这种酶，并克隆了人肽酶D的cDNA，确定了该酶的一级结构、基因的结构和基因的定位。我们对患者进行了基因分析。在一个有两个姐妹患有这种疾病的日本家庭中，我们发现培养细胞的 mRNA 存在 192 个碱基对 (bp) 偏转。我们从患者体内克隆了覆盖缺失突变的基因片段，并确定了核苷酸序列。结果表明，患者基因中大约有 759 bp 的序列被删除。在来自比利时的案例中（最初来自 Meddle East），我们发现了一个点突变（核苷酸 826 处的 A 到 G），导致氨基酸替换（Asp-Asn）。突变体 cDNA 的表达分析表明，无活性的片段是由 cDNA 合成的。另一位来自中东的患者也发现了同样的突变。因此，点突变可能起源于中东。此外，我们还确定了内含子-外显子连接周围的核苷酸序列。这些研究使我们能够扩增所有外显子以用于该疾病的基因分析。