Flow enhanced protein crystallization at the air/water interface

空气/水界面处的流动增强蛋白质结晶

• 批准号: 0755968
• 负责人: Amir Hirsa
• 金额: --
• 依托单位: Rensselaer Polytechnic Institute
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0755968&HistoricalAwards=false
• 关键词: Flow; enhanced; protein; crystallization; air

CBET-0755968HirsaThe ability to describe and use protein structure has enabled the rational design of new drugs and pharmacological agents. Many advances in understanding of biological systems at the molecular level have been made possible through detailed structural studies of proteins and nucleic acids. However, proteins must first be crystallized to use techniques that yield a precise description of their structure. In contrast to growing protein crystals in the bulk, 2D protein crystallization greatly simplifies both the theoretical and experimental aspects of protein studies. For example, 2D systems are not affected by gravity, an issue that plagues crystallographers with no affordable remedy. Two-dimensional protein crystallization at the air/water interface entails the specific binding of a protein to a lipid monolayer containing a ligand. The protein streptavidin, in solution, has been successfully crystallized at the air/water interface where a lipid monolayer containing a ligand, biotin, has been spread. The strong affinity between streptavidin and biotin initiates crystal formation at the interface under suitable conditions. The PIs will investigate the effects of flow in a system consisting of a stationary open cylinder driven by the constant rotation of the floor, in the axisymmetric flow regime over a wide range of Reynolds numbers. The PIs have recently discovered interface s sheared by flow can induce crystallization under conditions where none would otherwise occur. The PIs have discovered that flow can also produce a new form of crystal where the protein molecules appear to pack in a linear array, analogous to nano-wires, but much longer and perfectly straight. The PIs plan to investigate the macroscale flow phenomena associated with these new crystals. The PIs expect that varying the Reynolds number will regulate the nucleation rate and growth separately, which is a challenge for crystallographers. There are a vast number of proteins that have not been successfully crystallized by conventional means, and if flow-induced crystallization can improve the yield, then the effort here would be a resounding success. The multidisciplinary senior team (from mechanical engineering and applied mathematics), with its strong record of productive collaboration, will provide an excellent opportunity to educate graduate and undergraduate students in an exciting emerging field coupling hydrodynamics and protein studies.

CBET-0755968Hirsa描述和使用蛋白质结构的能力使得新药和药理制剂的合理设计成为可能。通过对蛋白质和核酸的详细结构研究，在分子水平上理解生物系统方面取得了许多进展。然而，蛋白质必须首先结晶，才能使用对其结构进行精确描述的技术。与大量生长蛋白质晶体相比，二维蛋白质结晶极大地简化了蛋白质研究的理论和实验方面。例如，二维系统不受重力影响，这个问题困扰着晶体学家，且没有经济实惠的解决方案。空气/水界面处的二维蛋白质结晶需要蛋白质与含有配体的脂质单层的特异性结合。溶液中的蛋白质链霉亲和素已在空气/水界面成功结晶，其中含有配体生物素的脂质单层已扩散。链霉亲和素和生物素之间的强亲和力在合适的条件下在界面处引发晶体形成。 PI 将研究由底板不断旋转驱动的固定开式圆柱体组成的系统中的流动效应，该系统在大范围雷诺数的轴对称流动状态下。 PI 最近发现，流动剪切的界面可以在原本不会发生的条件下诱导结晶。 PI 发现，流动还可以产生一种新形式的晶体，其中蛋白质分子似乎排列成线性阵列，类似于纳米线，但更长且完全笔直。 PI 计划研究与这些新晶体相关的宏观流动现象。 PI 预计改变雷诺数将分别调节成核速率和生长，这对晶体学家来说是一个挑战。有大量蛋白质尚未通过常规方法成功结晶，如果流动诱导结晶可以提高产量，那么这里的努力将取得巨大成功。多学科高级团队（来自机械工程和应用数学）凭借其良好的富有成效的合作记录，将为在流体动力学和蛋白质研究耦合的令人兴奋的新兴领域中教育研究生和本科生提供绝佳的机会。