In vitro study on the interaction between the 32 kDa enamelin and amelogenin.
In vitro study on the interaction between the 32 kDa enamelin and amelogenin.
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DOI：
10.1016/j.jsb.2009.01.003
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发表时间：
2009-04
影响因子：
3
通讯作者：
Moradian-Oldak J
中科院分区：
文献类型：
--
作者：
Fan D;Du C;Sun Z;Lakshminarayanan R;Moradian-Oldak J
釉质细胞外基质成分在控制釉质形成过程中晶体的成核和生长中起着至关重要的作用。我们研究了32 kDa的釉蛋白片段和釉原蛋白之间的相互作用，使用免疫化学和生物物理方法。免疫沉淀研究表明，32 kDa的釉蛋白和釉原蛋白一起从蛋白A柱洗脱。动态光散射结果表明，32 kDa的釉蛋白在pH 8.0的釉原蛋白组装有深远的影响，导致部分解离的纳米球，在剂量依赖性的方式。在添加32 kDa的釉蛋白后，在釉蛋白的圆二色性光谱中出现了一个等色性点，并且椭圆率最小值发生了偏移和强度降低，这表明釉蛋白的构象发生了变化，并且这两种大分子之间存在着直接的相互作用。我们的研究结果共同表明，32 kDa的釉蛋白在体外与釉原蛋白有直接的相互作用。我们目前的研究提供了新的见解，了解釉蛋白和釉原蛋白在大分子自组装和控制釉质矿物质的形成可能的合作。
Enamel extracelluar matrix components play vital roles in controlling crystal nucleation and growth during enamel formation. We investigated the interaction between the 32 kDa enamelin fragment and amelogenin using immunochemical and biophysical methods. Immunoprecipitation studies revealed that the 32 kDa enamelin and amelogenin eluted together from a Protein A column. Dynamic light scattering results showed that the 32 kDa enamelin had a profound effect on amelogenin assembly at pH 8.0, causing partial dissociation of the nanospheres, in a dose-dependent manner. The appearance of an isodichroic point and the shifting and intensity decrease of the ellipticity minima in the circular dichroism spectra of amelogenin following the addition of the 32 kDa enamelin were indicative of conformational changes in amelogenin and of a direct interaction between the two macromolecules. Our results collectively demonstrate that the 32 kDa enamelin has a direct interaction with amelogenin in vitro. Our current studies provide novel insights into understanding possible cooperation between enamelin and amelogenin in macromolecular self-assembly and in controlling enamel mineral formation.