Paine Michael L, Zhu Dan-Hong, Luo Wen, Snead Malcolm L
Center for Craniofacial Molecular Biology, School of Dentistry, University of Southern California, Los Angeles, Calif., USA.
Cells Tissues Organs. 2004;176(1-3):7-16. doi: 10.1159/000075023.
The secreted, full-length amelogenin is the dominant protein of the forming enamel organ. As enamel mineralization progresses, amelogenin is quickly subjected to proteolytic activity, and eliminated from the enamel environment. Mature enamel contains only traces of structural proteins, including enamelin and the sheath protein ameloblastin. In addition, a proteolytic fragment of amelogenin, known as the tyrosine-rich amelogenin peptide or TRAP, is present in low but isolatable quantities. By overexpressing TRAP during enamel development we sought to determine if such overexpression would result in structural alterations to the mature enamel. We reasoned that overexpressing a protein associated with enamel maturation, at an inappropriate developmental stage, would result in alterations to the enamel protein assembly and hence, alterations in enamel structure and morphology. As judged by transmission and scanning electron microscopy, the enamel formed by overexpressing TRAP showed little morphological differences when compared to the enamel of normal nontransgenic animals. Based on scanning electron-microscopic images, there was modest hypomineralization evident in the interrod enamel of the TRAP-overexpressing animals. However, this finding was inconsistent and inconsequential from a structural and functional perspective. From these results it appears that additional amounts of TRAP protein in the immature enamel matrix are not sufficient to alter the properties of the enamel extracellular matrix to an extent that the hierarchical structure of mature enamel is altered.
分泌型全长釉原蛋白是正在形成的釉质器官中的主要蛋白质。随着釉质矿化的进展,釉原蛋白很快受到蛋白水解作用,并从釉质环境中清除。成熟釉质仅含有痕量的结构蛋白,包括釉蛋白和鞘蛋白成釉蛋白。此外,釉原蛋白的一种蛋白水解片段,即富含酪氨酸的釉原蛋白肽(TRAP),含量虽低但可分离得到。在釉质发育过程中过表达TRAP,我们试图确定这种过表达是否会导致成熟釉质的结构改变。我们推断,在不适当的发育阶段过表达一种与釉质成熟相关的蛋白质,会导致釉质蛋白组装改变,从而导致釉质结构和形态改变。通过透射电子显微镜和扫描电子显微镜判断,与正常非转基因动物的釉质相比,过表达TRAP形成的釉质在形态上几乎没有差异。基于扫描电子显微镜图像,过表达TRAP的动物的釉柱间釉质有明显的轻度矿化不足。然而,从结构和功能的角度来看,这一发现并不一致且无关紧要。从这些结果看来,未成熟釉质基质中额外的TRAP蛋白量不足以改变釉质细胞外基质的性质,从而改变成熟釉质的层次结构。