Protein-mineral interactions at the organic-inorganic interface in biominerals

生物矿物质中有机-无机界面的蛋白质-矿物质相互作用

• 批准号: RGPIN-2016-05031
• 负责人: McKee, Marc
• 金额: $ 2.4万
• 依托单位: McGill University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2016
• 资助国家: 加拿大
• 起止时间: 2016-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=615454
• 关键词: Protein; mineral; interactions; organic; inorganic

Bones, teeth, otoconia, eggshells, snail and sea shells, corals and many other biomineralized structures in the plant and animal kingdoms arise from synergistic interactions between co-existing organic (usually proteins) and inorganic mineral phases. These composites have specialized properties, and hierarchically organized supramolecular assemblies that provide a framework for biomineralization. Negatively charged proteins may attain regulatory chemical complementarity by binding to mineral calcium at the organic-inorganic interface − a mechanism to influence crystal growth processes. It is hypothesized that the molecular precision of such organic-inorganic interfacial interactions regulates crystal growth. My biomineralization research program focuses on specific proteins/peptides (notably osteopontin) that regulate mineral growth. Intriguingly, amino acids, peptides and full-length proteins can be occluded within mineral crystals. Related to this, biomineralization can proceed initially through assembly of amorphous precursor nanoparticles forming within a confined, protein/peptide-rich reaction nanoenvironment. Within these nanoparticle domains, transformation towards a crystalline phase may occur over different length scales such that single crystals (by diffraction) can actually consist of aligned mineral nanoparticles, the fusion of which builds mesocrystals having occluded organics. This notion is in stark contrast to classical crystallization theory which postulates ion-by-ion attachment. My biomineralization research program compares these scenarios by exploring fundamental principles of how organics (amino acids, and relevant peptides and proteins) influence biomineralization. We will compare two polymorphs of calcium carbonate crystals (calcite and vaterite) grown in the presence of osteopontin protein/peptides/amino acids to two biomineralized structures − avian eggshell and mouse inner ear otoconia. To study the growth of calcium carbonate crystals in the presence of these organics, a variety of morphological, biochemical, immunochemical, cell biological/molecular, and characterization techniques will be used including: electron microscopy, atomic force microscopy, confocal microscopy, X-ray and electron diffraction, Canadian Light Source synchrotron analyses, Raman spectroscopy, immunocytochemistry on mouse otoconia, in vitro cell culture and crystal growth systems, mass spectroscopy, and RosettaSurface energy-minimization computational simulations. With this mechanistic biomineralization information on how proteins and peptides bind to crystals to regulate their growth, we will be well-positioned to create tunable mineralization events that advance biomaterials and tissue engineering applications to the benefit of Canadians and citizens worldwide.

骨骼，牙齿，牙齿，蛋壳，蜗牛和贝壳，珊瑚和许多其他生物矿化结构以及动物界是由共存有机有机（通常蛋白质）和无机矿物相之间的协同相互作用引起的。这些公司具有专门的物业，并组织了层次的超分子组件，可为生物矿化提供框架。带负电荷的蛋白质可能通过在有机无机界面上与矿物钙结合 - 一种影响晶体生长过程的机制来实现调节化学互补。假设这种有机无机相互作用的分子精度调节了晶体的生长。我的生物矿化研究计划的重点是调节矿物质生长的特定蛋白质/肽（尤其是骨桥）。 有趣的是，氨基酸，胡椒粉和全长蛋白可以在矿物质晶体中遮挡。与此相关的是，生物矿化最初可以通过组装在受约束的蛋白质/肽富反应纳米环境中形成的无定形前体纳米颗粒。在这些纳米颗粒结构域中，向晶相的转化可能会在不同的长度尺度上发生，因此单晶（通过衍射）实际上可以由对齐的矿物纳米颗粒组成，其融合构建具有闭塞组织的中晶的融合。这个概念与鉴定逐离子依恋的经典结晶理论形成鲜明对比。我的生物矿化研究计划通过探讨组织（氨基酸以及相关的胡椒剂和蛋白质）影响生物矿化的基本原理来比较这些情况。 我们将在存在骨桥蛋白蛋白/肽/氨基酸的情况下生长的两种多晶型碳酸钙晶体（方解石和阴道）与两个生物矿化结构-Avian Eggshell和小鼠内耳耳鼻喉科。为了研究这些组织存在的碳酸钙晶体的生长，将使用多种形态学，生化，免疫化学，细胞生物学/分子和特征技术，包括：电子显微镜，原子力显微镜，X射线显微镜，X射线显微镜，X射线和电子散射，Canadian Lightron分析，RAM RAM，RAM SYONERS，RAM SYONES同步，同步，同步，同步，同步，同步，同步，同步，小鼠耳鼻喉科，体外细胞培养和晶体生长系统，质谱和rosettasurface能量最小化计算模拟的免疫细胞化学。 借助有关蛋白质和辣椒如何与晶体结合以调节其生长的这种机械生物矿化信息，我们将有充分的位置，以创建可调的矿化事件，从而推动生物材料和组织工程的应用，以使全球加拿大人和公民的利益。