HIGH PRESSURE EXPERIMENTS WITH CUBIC INSULIN AND RHOMBOHEDRAL INSULIN

立方胰岛素和菱面体胰岛素的高压实验

• 批准号: 7358946
• 负责人: ALEXEI SUAREZ SOARES
• 金额: $ 0.45万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7358946
• 关键词: PRESSURE; EXPERIMENTS; CUBIC; INSULIN; RHOMBOHEDRAL

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. We are determining the effectiveness of using high pressure freezing of crystals to provide cryo protection, without the need for conventional chemical cryo protection. This evaluation will be useful in deciding how we with to proceed with respect to transfering this technology to the PxRR and the NSLS for use by the general user community. We are working in collaboration with the group at CHESS that developed this technology, and with the NSLS. Crystals of cubic insulin and rhombohedral insulin were prepared and frozen under vaying conditions of pressure and cryo protectant concentration. This method has previously proven most useful in crystals with relatively low solvent content, and the very large difference in the solvent content of the two specimen types (30% for rhobohedral insulin and 70% for cubic insulin) should provide data on this trend. A secondary objective of this project is to evaluate the density of the amorphous ice in the solvent region. The 011 reflection of cubic insulin is highly sensitive to the density contrast in the specimen, and we will track its intensity as the pressure is raised.

该子项目是利用 NIH/NCRR 资助的中心拨款提供的资源的众多研究子项目之一。子项目和研究者 (PI) 可能已从另一个 NIH 来源获得主要资金，因此可以在其他 CRISP 条目中得到体现。列出的机构是中心的机构，不一定是研究者的机构。我们正在确定使用晶体高压冷冻来提供低温保护的有效性，而不需要传统的化学低温保护。该评估将有助于决定我们如何将该技术转移到 PxRR 和 NSLS 以供一般用户社区使用。我们正在与开发这项技术的 CHESS 团队以及 NSLS 合作。制备立方胰岛素和菱形胰岛素晶体，并在不同的压力和冷冻保护剂浓度条件下冷冻。先前已证明该方法对于溶剂含量相对较低的晶体最有用，并且两种样品类型的溶剂含量差异非常大（菱形胰岛素为 30%，立方胰岛素为 70%）应该提供有关这一趋势的数据。 该项目的第二个目标是评估溶剂区域中无定形冰的密度。立方胰岛素的 011 反射对样本中的密度对比高度敏感，我们将随着压力升高跟踪其强度。