COMPOSITION OF HIGH DENSITY LIPOPROTEIN SUBCLASSES

高密度脂蛋白亚类的组成

• 批准号: 7724168
• 负责人: JOHN P KANE
• 金额: $ 0.5万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7724168
• 关键词: Age-Years; Antiatherogenic; Antibodies; Antibody Specificity; Apolipoprotein E; Atherosclerosis; Blindness; Cells; Cholesterol; Computer Retrieval of Information on Scientific Projects Database; Coronary Arteriosclerosis; Funding; Grant; High Density Lipoproteins; Human; Indium; Individual; Institution; Laboratories; Lipids; Lipoproteins; Macular degeneration; Mass Spectrum Analysis; Mediating; Metabolic; Metabolism; Methods; Molecular; Plasma; Play; Protease Inhibitor; Proteins; Research; Research Personnel; Resources; Retinal; Role; Source; Structure of retinal pigment epithelium; Transfer Factor; Transferase; United States National Institutes of Health; Western Blotting; hepatic lipase; insight; lipid metabolism; lipid transport; particle; phosphatidylcholine transfer protein

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. Our studies of lipoprotein metabolism, in particular, the high density lipoproteins (HDL) offer unique insights into lipid metabolism and transport that will be of value in understanding atherosclerosis at a molecular level. Methods developed in our laboratory have allowed us to document previously unreported molecular subspecies of HDL, each of which may play a specific antiatherogenic role associated with HDL. HDL subspecies purified from plasmas of normo- and dyslipidemic subjects reveal a variety of proteins. To date we have identified fifty-three candidate proteins that associate with discrete HDL particles. Among these , we have also discovered a new protein, designated apoL-1, that is associated with two discrete HDL species. While protein compositions of some HDL subspecies have been identified by Western blotting, we seek to obtain unequivocal characterization by mass spectroscopy especially in cases were our antibodies produce questionable identities. This frequently results from poor antibody specificity and recognition. Mass spectroscopic identification is of most benefit for the identification of proteins for which we have no specific antibodies, for proteins of low concentration, and for proteins which are not generally considered to associate with HDL. Our understanding of the HDL protein components consisting of lipid transfer factors such as cholesterol transfer protein, lecithin:cholesterol transferase, phosholipid transfer protein, hepatic lipase, plasma protease inhibitors, apoE, apoAIV, and apoL is paramount to our ability to understand the metabolic function of HDL-mediated protection in coronary artery disease. We have now begun to study the role of lipid transport in the retinal pigment epithelial cells, because they are key elements in human macular degeneration, the major cause of blindness in individuals over fifty years of age. This involves the identification of proteins expressed by the retinal cells, using mass spectrometry.

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 我们对脂蛋白代谢的研究，特别是高密度脂蛋白，提供了对脂代谢和运输的独特见解，这将有助于在分子水平上理解动脉粥样硬化。我们实验室开发的方法使我们能够记录以前未报道的高密度脂蛋白分子亚类，每个亚类都可能发挥与高密度脂蛋白相关的特定抗动脉粥样硬化作用。从正常和血脂异常受试者的血浆中提纯的高密度脂蛋白亚型揭示了多种蛋白质。到目前为止，我们已经确定了53个与离散的高密度脂蛋白颗粒相关的候选蛋白质。其中，我们还发现了一种新的蛋白质，命名为APOL-1，它与两个离散的高密度脂蛋白物种相关。虽然一些高密度脂蛋白亚种的蛋白质组成已经通过Western blotting确定，但我们寻求通过质谱学获得明确的表征，特别是在我们的抗体产生可疑身份的情况下。这通常是由于抗体的特异性和识别性较差造成的。对于我们没有特异性抗体的蛋白质、低浓度蛋白质以及通常认为与高密度脂蛋白无关的蛋白质的鉴定，质谱学鉴定是最有利的。我们对包括胆固醇转移蛋白、卵磷脂：胆固醇转移酶、磷脂转移蛋白、肝脂酶、血浆蛋白水解酶抑制物、apoE、apoAIV和APOL等脂转移因子的高密度脂蛋白组成的了解，对于我们理解高密度脂蛋白介导的冠状动脉疾病保护的代谢功能至关重要。 我们现在已经开始研究视网膜色素上皮细胞中脂质运输的作用，因为它们是人类黄斑变性的关键因素，黄斑变性是50岁以上人群失明的主要原因。这涉及到使用质谱学来鉴定视网膜细胞表达的蛋白质。