Structural, spatial, and temporal features guiding amelogenins transformation of calcium phosphate into enamel

引导牙釉蛋白将磷酸钙转化为牙釉质的结构、空间和时间特征

• 批准号: 10681277
• 负责人: Wendy J Shaw
• 金额: $ 48.76万
• 依托单位: BATTELLE PACIFIC NORTHWEST LABORATORIES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-19 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10681277
• 关键词: 3-Dimensional; Adsorption; Affect; Amelogenesis Imperfecta; Amino Acid Substitution; Amino Acids; Atomic Force Microscopy; Binding; Coupled; Crystallization; Dental Enamel; Development; Disease; Enamel Formation; Extracellular Matrix; Goals; Growth; Hydroxyapatites; In Situ; In Vitro; Individual; Ions; Knockout Mice; Knowledge; Maps; Microscopic; Minerals; Modification; Molecular; Morphology; Mus; Mutation; N-terminal; Nanosphere; Natural regeneration; Nature; Needles; Outcome; Persons; Phase; Phase Transition; Phenotype; Process; Property; Proteins; Raman Spectrum Analysis; Research; Roentgen Rays; Role; Scanning; Site; Spatial Distribution; Spectrum Analysis; Structure; System; Techniques; Therapeutic; Time; Tissues; Variant; Vertebrates; Work; X ray microscopy; amelogenin; biomineralization; calcium phosphate; design; falls; in vivo; insight; malformation; mineralization; mouse model; nanoscale; particle; prevent; protein distribution; protein protein interaction; protein structure; repaired; solid state nuclear magnetic resonance; spatial relationship; tomography; tool; transmission process

Project Summary The goal of the proposed work is to develop a molecular level, structural understanding of the role of amelogenin in directing the hierarchical growth of enamel. Enamel is one of the hardest biominerals known, and is hierarchically structured to prevent crack propagation, allowing it to last many people a lifetime. However, naturally occurring diseases or damage do occur to enamel and our current therapies fall far short of nature made enamel. Over the past decade we have come to understand that amelogenin assembles into quaternary structures as oligomers containing 12 proteins, units which further assemble into nanospheres. These assemblies, the molecular level interactions governing them, their interactions with calcium phosphate, and their role in mineral phase transformations, are not well understood for enamel formation. We do know that the protein amelogenin, the predominant protein in the forming enamel milieu, is critical in formation based on amelogenin null mouse models showing severely damaged enamel phenotypes. Protein structure often dictates function, yet we have only begun to reveal the structure of amelogenin. Further, the mineral in the enamel milieu begins as ions, and transitions through an amorphous stage prior to becoming elongated, crystallized HAP needles. Both in vivo and in vitro studies have demonstrated that amelogenin controls this phase transformation process, but the structural, spatial, and temporal aspects of this process are not understood. In our proposed work, we bring a set of advanced characterization techniques to an in vitro system simulating developing enamel to evaluate the structure of amelogenin when interacting with calcium phosphates, to understand the spatial distribution of amelogenin around developing calcium phosphate mineral ribbons, and to develop a molecular level view of the transformation process from calcium phosphate clusters, through the amorphous phase, on to crystalline HAP. This suite of techniques has never been brought to bear on understanding the challenge of enamel growth and will result in an understanding of the development of enamel that is currently lacking. The results of this work are relevant to biomineralization systems in general and may ultimately inform more robust therapeutics for enamel replacements.

项目概要 拟议工作的目标是建立对分子水平、结构的理解 釉原蛋白指导牙釉质的分层生长。牙釉质是已知最坚硬的生物矿物质之一， 其分层结构可防止裂纹扩展，使其能够陪伴许多人终生。 然而，牙釉质确实会发生自然发生的疾病或损伤，而我们目前的治疗方法远远达不到 大自然制造了珐琅。在过去的十年中，我们逐渐了解到釉原蛋白组装成 四级结构为含有 12 种蛋白质的寡聚物，这些单元进一步组装成纳米球。 这些组件，控制它们的分子水平相互作用，它们与磷酸钙的相互作用， 以及它们在矿物相变中的作用对于牙釉质的形成尚不清楚。我们确实知道 牙釉质蛋白是形成牙釉质环境中的主要蛋白质，对于牙釉质的形成至关重要 基于显示牙釉质表型严重受损的牙釉质缺失小鼠模型。蛋白质结构 通常决定了功能，但我们才刚刚开始揭示牙釉蛋白的结构。此外，矿物质中 牙釉质环境从离子开始，在变长之前经过非晶态阶段， 结晶 HAP 针。体内和体外研究都表明牙釉蛋白控制着这一点 相变过程，但该过程的结构、空间和时间方面并不 明白了。在我们提出的工作中，我们将一套先进的表征技术引入体外系统 模拟牙釉质发育以评估釉原蛋白与钙相互作用时的结构 磷酸盐，了解正在发育的磷酸钙矿物质周围牙釉蛋白的空间分布 丝带，并开发磷酸钙簇转化过程的分子水平视图， 通过非晶相，到结晶 HAP。这套技术从未被应用过 了解牙釉质生长的挑战，并将导致对牙釉质发育的理解 目前所缺乏的牙釉质。这项工作的结果与一般的生物矿化系统相关 并可能最终为牙釉质替代品提供更强大的治疗方法。

项目成果

Phosphorylation and ionic strength alter the LRAP-HAP interface in the N-terminus.

磷酸化和离子强度改变 N 末端的 LRAP-HAP 界面。

• 发表时间: 2013-04-02
• 影响因子: 2.9
• 作者: Lu, Jun;Xu, Yimin Sharon;Shaw, Wendy J
• 通讯作者: Shaw, Wendy J

Resolving protein-mineral interfacial interactions during in vitro mineralization by atom probe tomography.

通过原子探针断层扫描解决体外矿化过程中蛋白质-矿物质界面相互作用。

• 发表时间: 2023-06
• 作者: Taylor, Sandra D;Tao, Jinhui;Shin, Yongsoon;Buchko, Garry W;Dohnalkova, Alice;Grimm, Jack;Tarasevich, Barbara J;Ginovska, Bojana;Shaw, Wendy J;Devaraj, Arun
• 通讯作者: Devaraj, Arun

Changes in the quaternary structure of amelogenin when adsorbed onto surfaces.

吸附到表面时牙釉蛋白四级结构发生变化。

• DOI: 10.1002/bip.21095
• 发表时间: 2009-02
• 期刊: Biopolymers
• 影响因子: 2.9
• 作者: Tarasevich, Barbara J.;Lea, Scott;Bernt, William;Engelhard, Mark H.;Shaw, Wendy J.
• 通讯作者: Shaw, Wendy J.

Controls of nature: Secondary, tertiary, and quaternary structure of the enamel protein amelogenin in solution and on hydroxyapatite.

自然控制：溶液中和羟基磷灰石上釉质蛋白釉原蛋白的二级、三级和四级结构。

• 发表时间: 2020
• 影响因子: 3
• 作者: Shaw, Wendy J;Tarasevich, Barbara J;Buchko, Garry W;Arachchige, Rajith M J;Burton, Sarah D
• 通讯作者: Burton, Sarah D

Residue-Specific Insights into the Intermolecular Protein-Protein Interfaces Driving Amelogenin Self-Assembly in Solution.

对驱动釉原蛋白在溶液中自组装的分子间蛋白质-蛋白质界面的残基特异性洞察。

• 发表时间: 2022-12-20
• 影响因子: 2.9
• 作者: Buchko, Garry W;Mergelsberg, Sebastian T;Tarasevich, Barbara J;Shaw, Wendy J
• 通讯作者: Shaw, Wendy J