New Vistas in Vitamin K Metabolism

维生素 K 代谢的新前景

基本信息

批准号：
8477918
负责人：
Allan Edward Rettie
金额：
$ 28.77万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-05 至 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8477918
关键词：
Adopted Affect Anabolism Anticoagulants Automobile Driving Biochemical Biochemical Reaction Biological Biological Assay Blood Clot Blood coagulation Cancer Patient Catabolism Cell Line Cell model Cells Chemicals Chronic Kidney Failure Cytochrome P450 Data Development Diet Dimethylallyltranstransferase Engineering Enzymatic Biochemistry Enzymes Event Extrahepatic Factor IX Fat-Soluble Vitamin Foundations Fracture Future Genes Genetic Genetic Variation Genotype Goals Hand Hemostatic function Hepatic Hepatocyte Homeostasis Human Hydroquinones Hydroxylation Individual Knowledge Label Laboratories Link Liver Mass Spectrum Analysis Measures Metabolic Metabolism Mixed Function Oxygenases Nutrient Oral Pathway interactions Patients Pharmacogenetics Physiological Population Process Production Proteins RNA Interference Recombinants Relative (related person)Research Research Proposals Risk Risk Factors Role Sampling Series Side Source Supplementation Systems Analysis Testing Therapeutic Tissues Transfection Tumor Suppressor Genes Variant Vascular calcification Vitamin K Vitamin K 1 Vitamin K 2 Vitamin K Deficiency Warfarin analog base bone health bone metabolism calcification cancer therapy carboxylate carboxylation cell growth regulation cellular engineering cofactor combat coronary artery calcification dietary constituent enzyme activity genetic variant human tissue hydroquinone in vivo inhibitor/antagonist insight interest men mutant novel public health relevance response soft tissue tool

项目摘要

DESCRIPTION (provided by applicant): The long-term aim of this research is to define the network of integrated metabolic events that underlie vitamin K anabolism and catabolism, in order to understand inter-subject variability in response to this essential dietary nutrient. In ths new application, we will establish the P450 enzymes, CYP4F2 and CYP4F11 as the key vitamin K hydroxylases that initiate catabolism of this essential dietary constituent. In addition, we will determine the role of the newly identified human UBDIA1 enzyme in the interconversion of vitamins K1 and K2. These efforts will establish the link between the core vitamin K cycle proteins and ancillary enzymes that modulate the availability of the hydroquinone cofactors that drive Gla protein production. Because there is a high degree of inter-individual variability in response to supplementation with vitamin K in highly vulnerable K-deficient populations (e.g. those suffering from chronic kidney disease, cancer patients), these studies are of high significance. The goals of this research proposal are three-fold, (i) to establish the relative roes of human CYP4F2 and CYP4F11 in the initiation of vitamin K catabolism, (ii) to establish the role and chemical mechanism of action of UBIAD1 in conversion of vitamin K 1 to K2 (MK-4), and (iii) to ascertain the influence of common genetic variation in each of these three genes on vitamin K homeostatis in human tissues and cellular models of increasing complexity, as a precursor to future in vivo studies. In preliminary studies, we have accomplished the important tasks of heterologous expression of each of the three target enzymes, and development of highly sensitive and specific tandem MS assays for analysis of vitamin K in a variety of biological matrices. With these tools in hand, we propose, in Aim1, to evaluate the roles of CYP4F2 and CYP4F11 in vitamin K catabolism. In Aim 2, we will express and purify human UBIAD1 to enable an unambiguous assessment of the enzyme's catalytic capability and chemical mechanism for conversion of vitamin K1 to K3. In Aim 3, we will use banked, genotyped human liver tissue and a novel engineered cell system for analysis of Gla protein production to determine the effect of common genetic variation in these newly described vitamin K cycle-associated genes on cellular vitamin K homeostasis.

描述（由申请人提供）：这项研究的长期目的是定义综合代谢事件的网络，该综合代谢事件是维生素K的代谢性和分解代谢，以了解对这种基本饮食营养的响应，以理解受试者间的可变性。在新应用中，我们将建立P450酶，CYP4F2和CYP4F11作为关键的维生素K羟基酶，启动这种基本饮食成分的分解代谢。此外，我们将确定新鉴定的人ubdia1酶在维生素K1和K2互续中的作用。这些努力将建立核心维生素K循环蛋白与辅助酶之间的联系，该酶调节了驱动GLA蛋白产生的氢喹酮辅因子的可用性。由于在高度脆弱的K缺陷型种群中补充维生素K（例如那些患有慢性肾脏疾病的患者，癌症患者）的响应响应于维生素K，因此具有很高的个体间变异性，因此这些研究具有很高的意义。该研究建议的目标是三倍，（i）建立人CYP4F2和CYP4F11在启动维生素K分解代谢中的相对RO，（II）建立UBIAD1在维生素K 1至K2转化K2（MK-4）中的作用和化学机制，以确定属于Vitam k2（MK-4）的影响，以及（iii），属于属性的属性，这些变异属于属性的属性，这些变异属于这些变异的属性，这些变异属于这些变化的影响。在人体组织和增加复杂性的细胞模型中，作为未来研究中未来的先驱。在初步研究中，我们完成了三种靶酶异源表达的重要任务，并开发了高度敏感的和特定的串联MS测定法，以分析各种生物学基质中的维生素K。借助这些工具，我们在AIM1中建议评估CYP4F2和CYP4F11在维生素K分解代谢中的作用。在AIM 2中，我们将表达和净化人UBIAD1，以实现对酶的催化能力和化学机制的明确评估，以将维生素K1转化为K3。在AIM 3中，我们将使用库存的基因分型人肝组织和一种新型的工程细胞系统来分析GLA蛋白的产生，以确定这些新描述的维生素K循环相关基因对细胞维生素K稳态的影响。