Identification of novel amelogenin-binding proteins by proteomics analysis.
Identification of novel amelogenin-binding proteins by proteomics analysis.
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DOI：
10.1371/journal.pone.0078129
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Nishimura F
中科院分区：
文献类型：
--
作者：
Fukuda T;Sanui T;Toyoda K;Tanaka U;Taketomi T;Uchiumi T;Nishimura F
Emdogain(牙釉质基质衍生品，EMD)在牙周病学中得到广泛的认可。它用于牙周外科，以再生牙骨质、牙周膜和牙槽骨。然而，牙周再生的确切分子机制仍不清楚。在这项研究中，我们研究了与釉原蛋白结合的蛋白，这些蛋白被认为在促进牙周组织再生方面发挥了关键作用。为了确定与成釉蛋白相互作用并参与成骨细胞激活的新分子，我们采用亲和层析和蛋白质组学分析相结合的方法对分离的成骨细胞裂解物进行分析。在SAOS-2细胞中，胞浆中的许多釉原蛋白相互作用蛋白主要是细胞骨架蛋白和几个热休克蛋白70(HSP70)家族的伴侣分子。另一方面，细胞抽提物膜组分和胞质组分中的釉原蛋白相互作用蛋白的蛋白质组学特征有很大的不同。它们主要是内质网(ER)相关蛋白，少量的线粒体蛋白和核蛋白。在已鉴定的釉原蛋白相互作用蛋白中，我们验证了釉原蛋白与葡萄糖调节蛋白78(GRP78/Bip)的生物相互作用，GRP78/Bip存在于细胞液和膜富集组分中。共聚焦共定位实验有力地提示GRP78/Bip可能是釉原蛋白受体的候选基因。用GRP78/Bip基因敲除分析进一步的生物学评价。在本研究范围内，釉原蛋白与GRP78/Bip的相互作用促进了SAOS-2细胞的增殖，而与成骨分化无关。尽管其他相互作用的生物学意义尚不清楚，但这些发现表明，釉原蛋白来源的成骨细胞激活的不同效应可能对理解釉原蛋白诱导牙周组织再生的细胞和分子基础具有潜在的临床意义。
Emdogain (enamel matrix derivative, EMD) is well recognized in periodontology. It is used in periodontal surgery to regenerate cementum, periodontal ligament, and alveolar bone. However, the precise molecular mechanisms underlying periodontal regeneration are still unclear. In this study, we investigated the proteins bound to amelogenin, which are suggested to play a pivotal role in promoting periodontal tissue regeneration. To identify new molecules that interact with amelogenin and are involved in osteoblast activation, we employed coupling affinity chromatography with proteomic analysis in fractionated SaOS-2 osteoblastic cell lysate. In SaOS-2 cells, many of the amelogenin-interacting proteins in the cytoplasm were mainly cytoskeletal proteins and several chaperone molecules of heat shock protein 70 (HSP70) family. On the other hand, the proteomic profiles of amelogenin-interacting proteins in the membrane fraction of the cell extracts were quite different from those of the cytosolic-fraction. They were mainly endoplasmic reticulum (ER)-associated proteins, with lesser quantities of mitochondrial proteins and nucleoprotein. Among the identified amelogenin-interacting proteins, we validated the biological interaction of amelogenin with glucose-regulated protein 78 (Grp78/Bip), which was identified in both cytosolic and membrane-enriched fractions. Confocal co-localization experiment strongly suggested that Grp78/Bip could be an amelogenin receptor candidate. Further biological evaluations were examined by Grp78/Bip knockdown analysis with and without amelogenin. Within the limits of the present study, the interaction of amelogenin with Grp78/Bip contributed to cell proliferation, rather than correlate with the osteogenic differentiation in SaOS-2 cells. Although the biological significance of other interactions are not yet explored, these findings suggest that the differential effects of amelogenin-derived osteoblast activation could be of potential clinical significance for understanding the cellular and molecular bases of amelogenin-induced periodontal tissue regeneration.
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影响因子：
5.3
作者：
Haze A;Taylor AL;Haegewald S;Leiser Y;Shay B;Rosenfeld E;Gruenbaum-Cohen Y;Dafni L;Zimmermann B;Heikinheimo K;Gibson CW;Fisher LW;Young MF;Blumenfeld A;Bernimoulin JP;Deutsch D
通讯作者：
Deutsch D
影响因子：
4.1
作者：
Amin, Harsh D.;Olsen, Irwin;Donos, Nikolaos
通讯作者：
Donos, Nikolaos
影响因子：
4.8
作者：
Hatakeyama, J;Sreenath, T;Kulkarni, AB
通讯作者：
Kulkarni, AB
影响因子：
2.5
作者：
CHEN, EH;PIDDINGTON, R;GIBSON, CW
通讯作者：
GIBSON, CW
影响因子：
14.9
作者：
Fukuda, T;Ashizuka, M;Uchiumi, T
通讯作者：
Uchiumi, T