Geng Shuhui, Lei Yaping, Snead Malcolm L
The University of Southern California, Herman Ostrow School of Dentistry of USC, Center for Craniofacial Molecular Biology, Los Angeles, CA 90033.
School of Life Science and Technology, ShanghaiTech University, Shanghai, China, 201210.
JOM (1989). 2021 Jun;73(6):1696-1704. doi: 10.1007/s11837-021-04687-x. Epub 2021 May 7.
Amelogenin is the most abundant matrix protein guiding hydroxyapatite formation in enamel, the durable bioceramic tissue that covers vertebrate teeth. Here, we sought to refine structure-function for an amelogenin domain based on data showing a 42 amino acid amelogenin-derived peptide (ADP7) mimicked formation of hydroxyapatite similar to that observed for the full-length mouse 180 amino acid protein. In mice, we used CRISPR-Cas9 to express only ADP7 by the native amelogenin promoter. Analysis revealed ADP7 messenger RNA expression in developing mouse teeth with the formation of a thin layer of enamel. , ADP7 peptide partially replaced the function of the full-length amelogenin protein and its several protein isoforms. Protein structure-function relationships identified through assays can be deployed in whole model animals using CRISPR-Cas9 to validate function of a minimal protein domain to be translated for clinical use as an enamel biomimetic.
釉原蛋白是引导羟基磷灰石在牙釉质中形成的最丰富的基质蛋白，牙釉质是覆盖脊椎动物牙齿的耐用生物陶瓷组织。在此，我们试图基于数据完善釉原蛋白结构域的结构-功能，这些数据显示一种42个氨基酸的釉原蛋白衍生肽（ADP7）模拟了羟基磷灰石的形成，类似于在全长180个氨基酸的小鼠蛋白中观察到的情况。在小鼠中，我们使用CRISPR-Cas9通过天然釉原蛋白启动子仅表达ADP7。分析显示ADP7信使核糖核酸在发育中的小鼠牙齿中表达，并形成了一层薄的牙釉质。此外，ADP7肽部分替代了全长釉原蛋白及其几种蛋白异构体的功能。通过实验确定的蛋白质结构-功能关系可利用CRISPR-Cas9在全模型动物中进行验证，以确认最小蛋白结构域的功能，该结构域有望被翻译用于临床，作为一种牙釉质仿生材料。
