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.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 ACERT 的主要活动之一是开发提高生物医学研究通量的技术。这项重要的活动旨在加强 ACERT 的核心研究和合作，同时也有利于通过开发更灵敏、从而显着提高生产力的脉冲 ESR 光谱仪来激活其服务和传播活动。鉴于提高灵敏度具有最高优先级，因此在样品制备部分中不应将这项任务与 PDS 光谱学的另一方面分开考虑，因为样品的良好质量对于实现高通量也至关重要。为了实现这一目标，我们正在当地建立一个小型但设备齐全的设施，适合对蛋白质进行各种生化操作。重点将放在膜蛋白的研究上，日常活动将解决有效的蛋白质自旋标记问题；重组为脂质体、纳米盘或 bicelles、透析、样品浓缩等。我们希望通过能够制备和研究新鲜的膜蛋白样品来提高我们的工作质量，因为它们通常非常脆弱并且不能容忍过度的操作，例如冷冻/解冻循环。它将能够根据特定的蛋白质开发最有效的自旋标记方案，从而帮助我们的合作者分析自旋标记和优化 PDS 研究样本的能力有限或缺乏。 ACERT 工作人员将提供 ESR 光谱（特别是脉冲偶极 ESR）样品制备方面的 ACERT 用户支持和培训。 我们相信，这样的设施将有助于我们生物医学研究的灵活性和优点，并将支持该中心的效率。 此外，这样的设施将使 ACERT 科学家能够通过现场提供荧光光谱和圆二色性等标准技术来常规应用与 ESR 研究互补的方法。这将加强我们现有的研究，并使我们能够快速实施新的想法，这将有利于合作项目、核心研究和服务活动。