Full length amelogenin binds to cell surface LAMP-1 on tooth root/periodontium associated cells.
Full length amelogenin binds to cell surface LAMP-1 on tooth root/periodontium associated cells.
复制标题
DOI：
10.1016/j.archoralbio.2010.03.009
复制
发表时间：
2010-06
影响因子：
3
通讯作者：
Somerman, Martha J.
中科院分区：
文献类型：
--
作者：
Zhang, Hai;Tompkins, Kevin;Garrigues, Jacques;Snead, Malcolm L.;Gibson, Carolyn W.;Somerman, Martha J.
溶酶体相关膜蛋白-1（Lysosome-associated membrane protein-1，LAMP-1）是釉原蛋白（amelogenin，amelogenin）的一种细胞表面受体，是一种富含亮氨酸的釉原蛋白肽（leucine-rich amelogenin peptide，LRAP）。然而，目前还不清楚LAMP-1是否是牙间充质细胞的釉原蛋白受体。本研究的目的是确定LAMP-1是否作为牙根/牙周组织相关间充质细胞上全长釉原蛋白的细胞表面结合位点。将鼠牙囊细胞和成牙骨质细胞（OCCM-30）培养2天，然后加入全长重组小鼠釉原蛋白rp（H）M180。进行剂量-反应（0 - 100 μg/ml）和时间过程（0 - 120分钟）试验，以使用免疫荧光显微镜确定活细胞表面结合的最佳条件。通过向培养基中加入Emdogain（1 mg/ml），进行竞争性结合测定以确定结合特异性。使用针对LAMP-1的抗体检测LAMP-1在细胞表面上的位置，并将该模式与细胞表面结合的釉原蛋白进行比较。釉原蛋白和细胞表面的LAMP-1的免疫共定位，以比较的数量和分布模式。使用50 μg/ml rp（H）M180 120分钟达到最大表面结合。这种结合是特异性的，如通过加入Emdogain的竞争性抑制（降低79%）所证明的。rp（H）M180的结合模式与牙囊细胞和成牙骨质细胞表面LAMP-1的分布相似。rp（H）M180和LAMP-1的高共定位系数（0.92）支持rp（H）M180与细胞表面LAMP-1结合。本研究的数据表明，LAMP-1可以作为牙囊细胞和成牙骨质细胞上的釉原蛋白的细胞表面结合位点。
Lysosome-associated membrane protein-1 (LAMP-1) has been suggested to be a cell surface receptor for a specific amelogenin isoform, leucine-rich amelogenin peptide or LRAP. However, it is unclear if LAMP-1 is an amelogenin receptor for dental mesenchymal cells. The goal of this study was to determine if LAMP-1 serves as a cell surface binding site for full length amelogenin on tooth root/periodontium associated mesenchymal cells. Murine dental follicle cells and cementoblasts (OCCM-30) were cultured for 2 days followed by addition of full length recombinant mouse amelogenin, rp(H)M180. Dose-response (0 to 100 μg/ml) and time course (0 to 120 minutes) assays were performed to determine the optimal conditions for live cell surface binding using immuno-fluorescent microscopy. A competitive binding assay was performed to determine binding specificity by adding Emdogain (1 mg/ml) to the media. An antibody against LAMP-1 was used to detect the location of LAMP-1 on the cell surface and the pattern was compared to cell surface bound amelogenin. Both amelogenin and cell surface LAMP-1 were immuno-co-localized to compare the amount and distribution pattern. Maximum surface binding was achieved with 50 μg/ml of rp(H)M180 for 120 minutes. This binding was specific as demonstrated by competitive inhibition (79% lower) with the addition of Emdogain. The binding pattern for rp(H)M180 was similar to the distribution of surface LAMP-1 on dental follicle cells and cementoblasts. The high co-localization coefficient (0.92) for rp(H)M180 and LAMP-1 supports rp(H)M180 binding to cell surface LAMP-1. The data from this study suggest that LAMP-1 can serve as a cell surface binding site for amelogenin on dental follicle cells and cementoblasts.
登录
查看更多内容
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：
6.8
作者：
Dunker, A. Keith;Silman, Israel;Sussman, Joel L.
通讯作者：
Sussman, Joel L.
影响因子：
4.3
作者：
Boabaid, F;Gibson, CW;Somerman, MJ
通讯作者：
Somerman, MJ
影响因子：
4.8
作者：
Hatakeyama, J;Sreenath, T;Kulkarni, AB
通讯作者：
Kulkarni, AB
影响因子：
4.1
作者：
FINCHAM, AG;BELCOURT, AB;COTHRAN, WC
通讯作者：
COTHRAN, WC