Molecular Micellar-Polyacrylamide Gel Electrophoresis (MoMi-PAGE)

分子胶束-聚丙烯酰胺凝胶电泳 (MoMi-PAGE)

• 批准号: 7920804
• 负责人: ISIAH M WARNER
• 金额: $ 17.26万
• 依托单位: LOUISIANA STATE UNIV A&M COL BATON ROUGE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-15 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7920804
• 关键词: Acrylamides; Affect; Architecture; Area; Arterial Fatty Streak; Arteries; Belief; Blood; Blood capillaries; Charge; Chemicals; Chromatography; Complex Mixtures; Data; Diagnosis; Disease; Early Diagnosis; Experimental Designs; Fluorescent Probes; Funding; Goals; Grant; Health; Heart; Human; Hydrophobicity; Individual; Laboratories; Lead; Micelles; Molecular; Polyacrylamide Gel Electrophoresis; Polymers; Procedures; Proteins; Reagent; Research; Sampling; Sodium Dodecyl Sulfate; Solutions; Solvents; Structure; Techniques; Technology; Time; Tubular formation; United States National Institutes of Health; base; biological systems; capillary; chiral molecule; design; gel electrophoresis; human disease; improved; interest; molecular recognition; novel; stem; success; surfactant

DESCRIPTION (provided by applicant): This application outlines research which focuses on the use of molecular micelles to replace sodium dodecyl sulfate (SDS) and other conventional micelles in poly- acrylamide gel electrophoresis. The overall hypothesis of this research stems from our belief that molecular micelles should provide better PAGE separation of proteins than SDS or other conventional micelles. This hypothesis is based upon previous separation strategies developed in our laboratory for separation of chiral molecules. In addition, we have preliminary data provided in this application which supports this hypothesis. Thus, the overall objective of the research described here is to examine better separation strategies for analyses of protein samples using molecular micellar-polyacrylamide gel electrophoresis (MoMi-PAGE). To achieve this objective, this application is divided into three specific aims. The first specific aim is to understand the parameters which contribute to PAGE separations using molecular micelles as alternative reagents to SDS and other conventional micelles which are frequently used in PAGE separations. This first specific aim builds upon expertise developed over the past several years in our laboratory. Under this specific aim, we will explore various strategies with molecular micelles of specifically designed architectures to optimize separations for different classes of proteins using PAGE separations. Our second specific aim is to understand the mechanisms of our separations at the molecular level since previous success with our separation research has been the result of treating our separation problems as a molecular recognition problem. To achieve this aim, we will use fluorescent probes and experimental design to understand these parameters which contribute to optimal separations. Our third specific aim is to apply the technology that we have developed to the analyses of proteins associated with the formation of atherosclerotic plaques and proteins associated with other biological systems of interest. This latter specific aim is mostly descriptive of the kinds of application that we will ultimately explore since it is unlikely that we would get to such applications during the proposed two-year time frame of this application. Finally, it is noted that our previous studies under NIH funding, as well as the individual expertise of the collaborators brought together for this research, enhances the chance for success of the overall goals outlined in this application. Protein separations are very important to understanding many diseases which affect human health. Therefore, it is important to develop better separation and identification procedures for analyses of proteins. The most widely used protein separation technique is called sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) or (SDS-PAGE), which employs SDS for solubilization and separation of proteins on an electric grid. We propose to develop new reagents, call molecular micelles, to replace SDS and other surfactants in order to improve PAGE separation of proteins.

描述（由申请人提供）：本申请概述了重点关注在聚丙烯酰胺凝胶电泳中使用分子胶束替代十二烷基硫酸钠（SDS）和其他常规胶束的研究。这项研究的总体假设源于我们的信念，即分子胶束应该比 SDS 或其他传统胶束提供更好的蛋白质 PAGE 分离。该假设基于我们实验室先前开发的用于分离手性分子的分离策略。此外，我们在本申请中提供了支持这一假设的初步数据。因此，本文所述研究的总体目标是研究使用分子胶束聚丙烯酰胺凝胶电泳 (MoMi-PAGE) 分析蛋白质样品的更好的分离策略。为了实现这一目标，该应用程序分为三个具体目标。第一个具体目标是了解使用分子胶束作为 SDS 和其他常用于 PAGE 分离的常规胶束的替代试剂进行 PAGE 分离的参数。第一个具体目标建立在我们实验室过去几年开发的专业知识的基础上。在此特定目标下，我们将探索采用专门设计结构的分子胶束的各种策略，以利用 PAGE 分离优化不同类别蛋白质的分离。我们的第二个具体目标是在分子水平上了解我们的分离机制，因为我们之前的分离研究的成功是将我们的分离问题视为分子识别问题的结果。为了实现这一目标，我们将使用荧光探针和实验设计来了解这些有助于最佳分离的参数。我们的第三个具体目标是将我们开发的技术应用于分析与动脉粥样硬化斑块形成相关的蛋白质以及与其他感兴趣的生物系统相关的蛋白质。后一个具体目标主要描述了我们最终将探索的应用程序类型，因为我们不太可能在该应用程序建议的两年时间范围内获得此类应用程序。最后，值得注意的是，我们之前在 NIH 资助下的研究，以及为这项研究而汇集的合作者的个人专业知识，提高了本申请中概述的总体目标的成功机会。 蛋白质分离对于了解影响人类健康的许多疾病非常重要。因此，开发更好的蛋白质分析分离和鉴定程序非常重要。最广泛使用的蛋白质分离技术称为十二烷基硫酸钠 (SDS) 聚丙烯酰胺凝胶电泳 (PAGE) 或 (SDS-PAGE)，它采用 SDS 在电网上溶解和分离蛋白质。我们建议开发新的试剂，称为分子胶束，以取代 SDS 和其他表面活性剂，以改善蛋白质的 PAGE 分离。

项目成果

Positive cooperative mechanistic binding of proteins at low concentrations: a comparison of poly (sodium N-undecanoyl sulfate) and sodium dodecyl sulfate.

低浓度下蛋白质的正协同机械结合：聚（N-十一酰硫酸钠）和十二烷基硫酸钠的比较。

• DOI: 10.1016/j.jcis.2011.07.044
• 发表时间: 2011
• 期刊: Journal of colloid and interface science
• 影响因子: 9.9
• 作者: Das,Susmita;Sylvain,MonicaR;Fernand,VivianE;Losso,JackN;El-Zahab,Bilal;Warner,IsiahM
• 通讯作者: Warner,IsiahM