Studies on the genetic analysis of a patient with protein C deficien

1例蛋白C缺乏症患者的基因分析研究

• 批准号: 04671433
• 负责人: IDO Masaru
• 金额: $ 1.34万
• 依托单位: Mie 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: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04671433/
• 关键词: Protein C deficiency; Gene analysis; Prenatal diagnocis; Congenital anomaly; Thrombosis

We report genetic abnormalities of protein C gene in a male infant who developed neonatal purpura fulminans. DNA-sequence analysis of all cxons in protein C gene in this family revealed two mutations the first abnormality, derived from the mother, was a deletion of one of four consccutive g at nucleotide number 10758 in exon IX which would result in a frame shift mutation and com ; letely change amino acid sequence from Gly381 in the carboxyl-terminal region of protein C.The second abnormality, derived from the father, was a single nucleotide mutation from G to A in the codon (GAG to AAG) at nucleotide number 2977 in exon III, which would result in a substitution of Lys for gamma-carboxyglutamic acid (Gla) 26. This change would be responsible for the reduced immunological protein C levels of the patient and the father, estimated by a monoclonal antibody which recognizes the Gla-domain in a Ca^<2+>-dependent manner (3.8% and 57%, respectively). Partially purified abnormal protein C from the father's plasma showed a normal amidolytic activity and a change in the electrophoretic mobility. We detected the above mutations in his family members using two methods ; one was a creation of new restriction enzyme sites using mutagenic primers and the other was single nucleotide primer extension. Both methods are rapid and useful for the diagnosis of prenatal protein C abnormalities.

我们报告一个罹患新生儿爆发性紫癍症的男婴，其蛋白C基因有遗传异常。对该家系所有外显子的DNA序列分析表明，该家系有两个突变：第一个异常来自母亲，是外显子IX第10758位核苷酸的四个连续g中的一个缺失，导致移码突变和comc;在蛋白C的羧基末端区域从Gly 381开始的氨基酸序列发生了明显的改变。第二个异常，来自父亲，是外显子III中核苷酸编号2977的密码子中从G到A的单核苷酸突变（GAG到AAG），这将导致γ-羧基谷氨酸（Gla）26被Lys取代。这种变化可能是患者和父亲免疫学蛋白C水平降低的原因，这是通过一种单克隆抗体来估计的，该抗体以Ca^2+依赖的方式识别Gla结构域（分别为3.8%和57%）。从父亲的血浆中部分纯化的异常蛋白C显示出正常的酰胺分解活性和电泳迁移率的变化。我们用两种方法检测了他的家族成员中的上述突变;一种是使用诱变引物创建新的限制性内切酶位点，另一种是单核苷酸引物延伸。这两种方法都是快速和有用的产前蛋白C异常的诊断。

项目成果

Masaru Ido, Michiaki Ohiwa, Tatsuya Hayashi, Junji Nishioka, Tsuyoshi Hatada, Yasuyuki Watanabe, Hideo Wada, Shigeru Shirakawa, and Koji Suzuki: "A Compound Heterozygous Protein C Deficiency with a Single Nucleotide G Deletion Encoding Gly-381 and Amino A

Masaru Ido、Michiaki Ohiwa、Tatsuya Hayashi、Junji Nishioka、Tsuyoshi Hatada、Yasuyuki Watanabe、Hideo Wada、Shigeru Shirakawa 和 Koji Suzuki：“编码 Gly-381 和氨基 A 的单核苷酸 G 缺失的复合杂合蛋白 C 缺乏症

Masaru Ido,et al.: "A Compound Heterozygous Protein C Deficiency with a Single Nucleotide G Deletion Encoding Gly-381 and an Amino Acid Substitution of Lys for Gla-26" Thrombosis and Haemostasis. 70. 636-641 (1993)

Masaru Ido 等人：“复合杂合蛋白 C 缺乏症，编码 Gly-381 的单核苷酸 G 缺失以及用 Lys 氨基酸取代 Gla-26”血栓形成和止血。