PATHOPHYSIOLOGY OF BILIRUBIN BOUND COVALENTLY TO ALBUMIN

胆红素与白蛋白共价结合的病理生理学

• 批准号: 3079005
• 负责人: ANIL GAUTAM
• 金额: $ 7.51万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-04-01 至 1987-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3079005
• 关键词: benzodiazepines; biliary tract disorder; bilirubin; bilirubin glucuronide; chemical structure; covalent bond; estrogens; high performance liquid chromatography; human tissue; hyperbilirubinemia; liver metabolism; molecular site; perfusion; thin layer chromatography

The major objective of this research project is to investigate the pathophysiological significance of BIL-ALB, recently confirmed by us to be a significant fraction of serum conjugated bilirubin (BR) that is apparently covalently bound to albumin is sera from patients with hepatobiliary diseases. Specific aims: To investigate the chemical nature and site of formation of BIL-ALB and to determine if other organic anions also bind covalently to albumin in the presence of cholestasis. Methods: BIL-ALB and other BR fractions will be measured by a reverse-phase HPLC method. BIL-ALB will be purified from pooled, jaundiced sera by sequential column chromatography. The identity of the conjugated BR species will be determined by thin-layer chromatography (TLC) of ethyl anthranilate azoderivatives and products of methanolysis with sodium methoxide. Proteolysis of BIL-ALB followed by purification and Edman-degradation of BR-tagged 'chromopeptides' will be used to identify the amino acid site/s of binding. Chromic acid oxidation and TLC separation of the liberated imides would aid in identifying the nature of the BR-albumin bond. An isolated perfused rat liver model will be used, along with detailed kinetic studies of any non-enzymatic formation of BIL-ALB in in vitro systems to determine the site of BIL-ALB synthesis- in the liver or in peripheral circulation. Any alteration in the turnover rate of albumin resulting from BIL-ALB formation will be studied. In addition, covalent binding of estrogen and benzodiazepene metabolites to albumin will be looked for in patients with cholestasis. Significance: These studies should better our understanding of the metabolism, transport and protein-binding of BR and other organic anions in the presence of hepatobiliary diseases and would lead to a more complete study of protein binding of organic anions. After completing 2 years of laboratory research supported by an NIH Institutional Training grant in Hepatology, the candidate was appointed to the Digestive Diseases Division faculty in July, 1983. The Yale Liver Study Unit is an ideal environment for this project. The co-sponsors are well experienced researchers in hepatocellular function and metabolism. Dr. E. Gordon is an internationally renowned expert in bilirubin chemistry and metabolism. Thus, both equipment and technical expertise are excellent. Interaction with basic science department such as Biochemistry and Chemistry is readily available

本研究项目的主要目的是调查 BIL-ALB的病理生理学意义，最近我们证实， 血清结合胆红素（BR）的显著部分， 与白蛋白明显共价结合的是来自患有 肝胆疾病 具体目的：研究化学性质 和BIL-ALB的形成部位，并确定其他有机阴离子是否 在胆汁淤积的情况下也与白蛋白共价结合。 研究方法： BIL-ALB和其他BR组分将通过反相HPLC进行测量 法 BIL-ALB将从合并的黄疸血清中纯化， 柱层析 共轭BR物质的特性将是 邻氨基苯甲酸乙酯薄层色谱法测定 偶氮衍生物和甲醇钠甲醇分解产物。 BIL-ALB的蛋白水解，随后的纯化和Edman降解 BR标记的“色肽”将用于鉴定氨基酸位点 的约束。 铬酸氧化和薄层色谱分离 酰亚胺将有助于鉴定BR-白蛋白键的性质。 一个 将使用离体灌流大鼠肝脏模型，沿着详细的动力学 体外系统中BIL-ALB的任何非酶促形成的研究， 确定BIL-ALB合成的位点-在肝脏或外周 流通 由以下原因引起的白蛋白周转率的任何改变 将研究BIL-ALB形成。 此外，共价结合 雌激素和苯二氮卓类代谢物与白蛋白的结合将在 胆汁淤积患者。 意义：这些研究应该更好地 了解BR的代谢、转运和蛋白结合， 其他有机阴离子在肝胆疾病的存在下， 导致对有机阴离子蛋白质结合的更完整的研究。 在完成了由NIH支持的2年实验室研究后， 在肝病学机构培训补助金，候选人被任命为 1983年7月，消化疾病科教员。 耶鲁肝脏 学习单元是这个项目的理想环境。 共同赞助者是 在肝细胞功能和代谢方面经验丰富的研究人员。 博士E.戈登是国际知名的胆红素化学专家 和新陈代谢。 因此，设备和技术专长都是 好极了 与生物化学等基础科学部门的互动 而化学是现成的

项目成果

Menopausal estrogen use, high density lipoprotein cholesterol subfractions and liver function.

更年期雌激素的使用、高密度脂蛋白胆固醇亚组分和肝功能。

• DOI: 10.1016/0021-9150(83)90005-9
• 发表时间: 1983
• 期刊: Atherosclerosis
• 影响因子: 5.3
• 作者: Cauley,JA;LaPorte,RE;Kuller,LH;Bates,M;Sandler,RB
• 通讯作者: Sandler,RB

Determinants of bone mass in menopause.

更年期骨量的决定因素。

• DOI: 10.1016/0091-7435(82)90053-6
• 发表时间: 1982
• 期刊: Preventive medicine
• 影响因子: 5.1
• 作者: Black-Sandler,R;LaPorte,RE;Sashin,D;Kuller,LH;Sternglass,E;Cauley,JA;Link,MM
• 通讯作者: Link,MM