Activation of extracellular signal-regulated kinases 1 and 2 (ERK1 and ERK2) by FGF-2 and PDGF-BB in normal human osteoblastic and bone marrow stromal cells: Differences in mobility and in-gel renaturation of ERK1 in human, rat, and mouse osteoblastic cells
Activation of extracellular signal-regulated kinases 1 and 2 (ERK1 and ERK2) by FGF-2 and PDGF-BB in normal human osteoblastic and bone marrow stromal cells: Differences in mobility and in-gel renaturation of ERK1 in human, rat, and mouse osteoblastic cells
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DOI：
10.1006/bbrc.1997.7256
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发表时间：
1997-09-08
影响因子：
3.1
通讯作者：
Avioli, LV
中科院分区：
文献类型：
--
作者：
Chaudhary, LR;Avioli, LV
我们最近证明，碱性成纤维细胞生长因子（FGF-2）和血小板衍生生长因子- bb （PDGF-BB）主要激活细胞外信号调节激酶2 （ERK2）在正常的人成骨细胞（NOB）和骨髓基质（HBMS）细胞中通过“凝胶内”MAP激酶测定，尽管ERK1和ERK2蛋白都存在。在本研究中，我们检测了ERK1是否也被生长因子激活，通过使用三种不同的MAPK检测程序，“凝胶内MAP激酶检测”，免疫复合物激酶检测，以及用抗活性MAPK抗体（识别ERK1和ERK2的特异性激活形式）进行western blotting。结果表明，除了ERK2， ERK1在正常的HOB和HEMS细胞中还被FGF-2和PDGF-BB激活。与大鼠和小鼠相比，人ERK1在sds -聚丙烯酰胺凝胶上移动，揭示了正常人成骨细胞和骨髓骨祖细胞、人（TE-85）和大鼠（ROS 17/2.8和UMR-106）骨肉瘤和小鼠（MC3T3-E1）成骨细胞中ERK1表观分子量的差异，ERK1在凝胶内再生过程中比ERK2更不稳定；因此，凝胶内MAP激酶检测不能准确估计ERK1的活性，结果也表明抗活性MAPK抗体可以可靠、准确地测量成骨细胞中ERK1和ERK2的活性。(C) 1997学术出版社。
We recently demonstrated that basic fibroblast growth factor (FGF-2) and platelet-derived growth factor-BB (PDGF-BB) mainly activated extracellular signal-regulated kinase 2 (ERK2) in normal human osteoblastic (NOB) and bone marrow stromal (HBMS) cells by are ''in-gel'' MAP kinase, assay, although both ERK1 and ERK2 proteins were present. In the present study, we examined whether ERK1 is also activated by growth factors by using three different MAPK assay procedures, an ''in-gel MAP kinase assay,'' an immune-complex kinase assay, and western blotting with anti-active MAPK antibody which recognizes specifically activated forms of both ERK1 and ERK2, Results have demonstrated that in addition to ERK2, ERK1 is activated by FGF-2 and PDGF-BB in normal HOB and HEMS cells, The human ERK1 moved Taster on SDS-polyacrylamide gel compared to rat and mouse, revealing differences in the apparent molecular weight of ERK1 in normal human osteoblastic and bone marrow osteoprogenitor cells, human (TE-85) and rat (ROS 17/2.8 and UMR-106) osteosarcoma, and mouse (MC3T3-E1) osteoblastic cells, ERK1 is less stable in the in-gel renaturation process compared to ERK2; thus, in-gel MAP kinase assay does not provide an accurate estimation of ERK1 activity, Results also showed that anti-active MAPK antibody can be used reliably and accurately to measure the activation of ERK1 and ERK2 in osteoblastic cells. (C) 1997 Academic Press.