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-0755968HIRSATHE描述和使用蛋白质结构的能力使新药和药理剂的合理设计能够。通过详细的蛋白质和核酸的结构研究，对分子水平生物系统的理解有许多进步。但是，必须首先将蛋白质结晶以使用对其结构进行精确描述的技术。与大量蛋白质晶体生长相反，2D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，2D系统不受重力的影响，这是一个困扰没有负担得起的疗法的晶体学家的问题。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。溶液中的蛋白链霉亲和蛋白已在空气/水界面上成功结晶，其中含有配体生物素的脂质单层已扩散。链霉亲和素之间的强亲和力在适当的条件下启动界面处的晶体形成。 PI将研究由固定的开放气缸组成的系统中流动的效果，该系统由地板的恒定旋转驱动，在轴对称流程中的恒定旋转范围内，在较大的雷诺数上。 PI最近发现了流动剪切的界面可以在不发生任何情况的情况下诱导结晶。 PI发现流动还可以产生一种新的晶体形式，其中蛋白质分子似乎包装在线性阵列中，类似于纳米系列，但更长，更完美。 PIS计划研究与这些新晶体相关的宏观流动现象。 PI期望改变雷诺数的数量将分别调节成核率和生长，这对晶体学家来说是一个挑战。有许多蛋白质尚未成功通过常规手段结晶，如果流动诱导的结晶可以提高产量，那么这里的努力将是巨大的成功。多学科的高级团队（来自机械工程和应用数学）凭借其富有生产力的合作记录，将提供一个极好的机会，以教育研究生和本科生在令人兴奋的新兴领域耦合流体动力学和蛋白质研究。