SELECTIVE SECRETION PATHWAY MEDIATED BY LMAN1 AND MCFD2

LMAN1 和 MCFD2 介导的选择性分泌途径

• 批准号: 7602906
• 负责人: David Ginsburg
• 金额: $ 2.33万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7602906
• 关键词: Accounting; Alleles; Complex; Computer Retrieval of Information on Scientific Projects Database; Endoplasmic Reticulum; Factor V Deficiency; Factor VIII; Family; Funding; Genes; Goals; Golgi Apparatus; Grant; Immunoprecipitation; Institution; Mediating; Messenger RNA; Missense Mutation; Mutation; Pathway Analysis; Pathway interactions; Patients; Plasma Proteins; Proteins; Reporting; Research; Research Personnel; Resources; Source; United States National Institutes of Health; Western Blotting; clinically significant; lymphoblast; receptor

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Mutations in LMAN1 (ERGIC-53) or MCFD2 cause combined deficiency of factor V and factor VIII (F5F8D). LMAN1 and MCFD2 form a protein complex that functions as a cargo receptor ferrying FV and FVIII from the endoplasmic reticulum to the Golgi. We analyzed 10 previously reported and 10 new F5F8D families. Mutations in the LMAN1 or MCFD2 genes accounted for 15 of these families, including 3 alleles resulting in no LMAN1 mRNA accumulation. Combined with our previous reports, we have identified LMAN1 or MCFD2 mutations as the causes of F5F8D in 71 of 76 families. Among the 5 families in which no mutations were identified, 3 were due to misdiagnosis, with the remaining 2 likely carrying LMAN1 or MCFD2 mutations that were missed by direct sequencing. Our results suggest that mutations in LMAN1 and MCFD2 may account for all cases of F5F8D. Immunoprecipitation and Western blot analysis detected a low level of LMAN1-MCFD2 complex in lymphoblasts derived from patients with missense mutations in LMAN1 (C475R) or MCFD2 (I136T), suggesting that complete loss of the complex may not be required for clinically significant reduction in FV and FVIII. The immediate goal of this project is to identify additional proteins that are secreted via this pathway by analysis of plasma protein levels.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 LMAN1 (ERGIC-53) 或 MCFD2 突变导致 V 因子和 VIII 因子 (F5F8D) 联合缺乏。 LMAN1 和 MCFD2 形成蛋白质复合物，充当货物受体，将 FV 和 FVIII 从内质网运送到高尔基体。我们分析了 10 个先前报告的 F5F8D 系列和 10 个新的 F5F8D 系列。其中 15 个家族发生了 LMAN1 或 MCFD2 基因突变，其中 3 个等位基因导致 LMAN1 mRNA 不积累。结合我们之前的报告，我们在 76 个家庭中的 71 个家庭中确定了 LMAN1 或 MCFD2 突变是 F5F8D 的原因。在未发现突变的5个家系中，有3个是由于误诊所致，其余2个可能携带LMAN1或MCFD2突变，但直接测序漏掉了这些突变。我们的结果表明 LMAN1 和 MCFD2 的突变可能是所有 F5F8D 病例的原因。免疫沉淀和蛋白质印迹分析检测到 LMAN1 (C475R) 或 MCFD2 (I136T) 错义突变患者的淋巴母细胞中存在低水平的 LMAN1-MCFD2 复合物，表明临床上显着降低 FV 可能不需要完全丧失该复合物和FVIII。 该项目的直接目标是通过分析血浆蛋白水平来识别通过该途径分泌的其他蛋白质。