CryoTEM study of effects of phosphorylation on the hierarchical assembly of porcine amelogenin and its regulation of mineralization in vitro.
CryoTEM study of effects of phosphorylation on the hierarchical assembly of porcine amelogenin and its regulation of mineralization in vitro.
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DOI：
10.1016/j.jsb.2013.05.011
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发表时间：
2013-08
影响因子：
3
通讯作者：
Beniash, Elia
中科院分区：
文献类型：
--
作者：
Fang, Ping-An;Margolis, Henry C.;Conway, James F.;Simmer, James P.;Beniash, Elia
关键词：
釉原蛋白是主要的细胞外牙釉质基质蛋白，在调节牙釉质的生长和组织中起着至关重要的作用。通过低温透射电子显微镜 (cryoTEM) 研究了全长天然 (P173) 和重组 (rP172) 猪牙釉蛋白的组装和矿化。冷冻透射电镜显示，天然和重组猪牙釉蛋白均经历逐步自组装。尽管 P173 和 rP172 寡聚物的整体结构组织相似并且类似于鼠重组牙釉蛋白 rM179 的寡聚物，但存在细微的差异，表明 P173 中存在的单个磷酸化丝氨酸可能会影响牙釉蛋白自组装。我们的矿化研究表明，P173 和 rP172 低聚物都能稳定初始矿物簇。然而，重要的是，rP172 调节初始矿物簇的组织成线性链，并引导细长矿物颗粒平行阵列的形成，这是牙釉质结构组织的标志。这些结果与之前使用全长重组鼠牙釉蛋白获得的结果相似。与 rP172 所见相反，磷酸化 P173 会在较长时间内强烈抑制矿化。我们认为这些差异可能是由于 P173 和 rP172 之间结构组织和电荷分布的差异造成的。总的来说，我们的研究表明牙釉蛋白的自组装及其控制矿化的机制可能在不同的哺乳动物物种中具有普遍性。我们的数据还为磷酸化对牙釉蛋白自组装的影响及其矿化调节提供了新的见解。
Amelogenin, the major extracellular enamel matrix protein, plays a critical role in regulating the growth and organization of enamel. Assembly and mineralization of full-length native (P173) and recombinant (rP172) porcine amelogenins were studied by cryogenic Transmission Electron Microscopy (cryoTEM). The cryoTEM revealed that both native and recombinant porcine amelogenins undergo step-wise self-assembly. Although the overall structural organization of P173 and rP172 oligomers was similar and resembled oligomers of murine recombinant amelogenin rM179, there were subtle differences suggesting that a single phosphorylated serine present in P173 might affect amelogenin self-assembly. Our mineralization studies demonstrated that both P173 and rP172 oligomers stabilize initial mineral clusters. Importantly, however, rP172 regulated the organization of initial mineral clusters into linear chains and guided the formation of parallel arrays of elongated mineral particles, which are the hallmark of enamel structural organization. These results are similar to those obtained previously using full-length recombinant murine amelogenin. In contrast to that seen with rP172, phosphorylated P173 strongly inhibits mineralization for extended periods of time. We propose that these differences might be due to the differences in the structural organization and charge distribution between P173 and rP172. Overall our studies indicate that self-assembly of amelogenin and the mechanisms of its control over mineralization might be universal across different mammalian species. Our data also provide new insight into the effect of phosphorylation on amelogenin self-assembly and its regulation of mineralization.
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影响因子：
7.6
作者：
Bouropoulos, N;Moradian-Oldak, J
通讯作者：
Moradian-Oldak, J
影响因子：
3.8
作者：
Iijima, Mayumi;Fan, Daming;Bromley, Keith M.;Sun, Zhi;Moradian-Oldak, Janet
通讯作者：
Moradian-Oldak, Janet
影响因子：
3
作者：
Beniash, Elia;Metzler, Rebecca A.;Lam, Raymond S. K.;Gilbert, P. U. P. A.
通讯作者：
Gilbert, P. U. P. A.
影响因子：
2.7
作者：
Hu, Jan C. -C.;Chun, Yong-Hee P.;Simmer, James P.
通讯作者：
Simmer, James P.
影响因子：
14
作者：
Bajaj, Devendra;Arola, Dwayne D.
通讯作者：
Arola, Dwayne D.