BUILDING UP THE FACILITY FOR MEMBRANE PROTEIN MANIPULATION AND SPIN LABELING

建立膜蛋白操作和旋转标记设施

• 批准号: 8172235
• 负责人: ELKA R GEORGIEVA
• 金额: $ 1.13万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8172235
• 关键词: Address; Back; Biochemical; Biological Assay; Biomedical Research; Circular Dichroism Spectroscopy; Collaborations; Computer Retrieval of Information on Scientific Projects Database; Dialysis procedure; Electron Spin Resonance Spectroscopy; Freezing; Funding; Goals; Grant; Institution; Liposomes; Membrane Proteins; Methods; Physiologic pulse; Preparation; Proteins; Protocols documentation; Research; Research Personnel; Resources; Sampling; Scientist; Services; Side; Site; Source; Spectrum Analysis; Spin Labels; Techniques; Technology; Training Support; United States National Institutes of Health; Work; flexibility; improved; reconstitution

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. One of the major activities at ACERT is aimed at developing technologies for increasing the throughput of biomedical research. This important activity is poised to strengthen ACERT core research and collaborations, but also to benefit its service and dissemination activates by means of developing more sensitive and thereby considerably more productive pulse ESR spectrometers. Given that increasing sensitivity carries the highest priority, this task should not be considered in isolation from the other side of PDS spectroscopy in the part of sample preparation, since sample good quality is also essential for achieving high throughput. To address this goal, we are setting up a small but well-equipped local facility, suited for variety of biochemical manipulation with proteins. The emphasis will be given to the research on membrane proteins and everyday activities will be addressing efficient protein spin labeling; reconstitutions into liposomes, nanodisks or bicelles, dialysis, sample concentration, etc. We expect to improve the quality of our work by having been able to prepare and study fresh membrane protein samples, since they often are very delicate and do not tolerate excessive manipulations, for example freezing/thawing cycles. It will enable developing the most efficient protocols for spin-labeling, depending on the particular protein, thus helping our collaborators who have limited or lacking capacity to assay spin-labeling and optimize samples for PDS study. ACERT user support and training in sample preparation for ESR spectroscopy, in particular for pulsed dipolar ESR will be provided by ACERT staff. We are confident that such facility will contribute to the flexibility and merits of our biomedical research and will back the efficiency of the center. Additionally, such a facility will enable ACERT scientists to apply routinely the methods complementary to the ESR studies by providing on site access to standard techniques, such as fluorescent spectroscopy and circular dichroism. This will strengthen our existing research, and enable us to quickly implement the new ideas, which will be beneficial for collaborative projects, core research, and service activities.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 ACERT的主要活动之一旨在开发用于增加生物医学研究吞吐量的技术。这项重要的活动有望加强ACERT核心研究和协作，但也通过开发更敏感的脉搏ESR ESR光谱仪而受益于其服务和传播会激活其服务和传播。鉴于提高敏感性具有最高优先级，因此在样品制备的一部分中，不应从PDS光谱的另一侧隔离此任务，因为样品质量良好对于实现高吞吐量也是必不可少的。为了解决这一目标，我们正在建立一个小型但设备齐全的本地设施，适用于使用蛋白质进行多种生化操作。将重点放在有关膜蛋白的研究中，日常活动将解决有效的蛋白质自旋标记。对脂质体，纳米虫或双丝，透析，样品浓度等的重建。我们希望能够通过能够准备和研究新鲜的膜蛋白样品来提高工作质量，因为它们通常非常精致，并且不耐受过多的操纵，例如冷冻/裂开/冻结的环境。它将根据特定的蛋白质制定最有效的自旋标记方案，从而帮助我们的合作者有限或缺乏能力分析自旋标记并优化PDS研究样本的能力。 ACERT用户支持和ESR光谱样品准备中的培训，尤其是脉冲双极ESR的ACERT人员将由ACERT员工提供。 我们相信，这种设施将有助于我们生物医学研究的灵活性和优点，并支持该中心的效率。 此外，这样的设施将使ACERT科学家通过在现场访问标准技术（例如荧光光谱和圆形二分法）方面定期应用与ESR研究的方法。这将加强我们的现有研究，并使我们能够快速实施新想法，这将对协作项目，核心研究和服务活动有益。