Modulation of bone resorption by phosphorylation state of bone sialoprotein.
Modulation of bone resorption by phosphorylation state of bone sialoprotein.
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DOI：
10.1021/bi900066b
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发表时间：
2009-07-28
期刊：
影响因子：
2.9
通讯作者：
Salih, Erdjan
中科院分区：
文献类型：
--
作者：
Curtin, Paul;McHugh, Kevin P.;Zhou, Hai-Yan;Fluckiger, Rudolf;Goldhaber, Paul;Oppenheim, Frank G.;Salih, Erdjan
我们确定了培养的破骨细胞和巨噬细胞中跨膜蛋白酪氨酸磷酸化（由外向内信号传导），以响应添加的天然纯化 BSP (nBSP) 及其去磷酸化形式 (dBSP)。 dBSP 对巨噬细胞和破骨细胞的细胞内酪氨酸磷酸化具有选择性/差异性和有效的抑制作用，而 nBSP 对细胞内酪氨酸磷酸化的影响最小。对下游基因表达效应的进一步研究导致在巨噬细胞和破骨细胞中鉴定出大量响应 nBSP 相对于 dBSP 的差异表达基因。这些研究扩展到骨吸收模型，使用活体小鼠新生儿颅骨骨器官培养物刺激甲状旁腺激素（PTH）进行骨吸收。在此类培养物中添加 nBSP 对 I 型胶原端肽片段释放没有影响，因此对整体骨吸收没有影响，而添加 dBSP 则消除了 PTH 诱导的骨吸收。 dBSP 对骨吸收的抑制作用被证明是独特的，因为在补充实验中，使用整合素受体结合配体 GRGDS 肽仅部分减少总体骨吸收。定量 RANKL 分析表明，dBSP 通过下调成骨细胞 RANKL 的产生来抑制 PTH 诱导的骨吸收。这一结论得到了培养的 MC3T3-E1 成骨细胞的 RANKL 分析的支持。总体而言，这些研究提供了直接证据，证明 BSP 上的共价结合磷酸盐通过跨膜酪氨酸磷酸化参与受体介导的“由外向内”信号传导，同时对下游基因表达产生影响。活骨器官培养系统的使用增强了这些结果，并提供了进一步的证据，将观察到的体内磷酸化可变状态与骨重塑联系起来。
We have determined transmembrane protein tyrosine phosphorylation (outside-in signaling) in cultured osteoclasts and macrophages in response to added native purified BSP (nBSP) and its dephosphorylated form (dBSP). There were selective/differential and potent inhibitory effects by dBSP and minimal effect by nBSP on intracellular tyrosine phosphorylation in macrophages and osteoclasts. Further studies on the down-stream gene expression effects led to identification of a large number of differentially expressed genes in response to nBSP relative to dBSP in both macrophages and osteoclasts. These studies were extended to bone resorption model using live mouse neonatal calvarial bone organ cultures stimulated by parathyroid hormone (PTH) to undergo bone resorption. Inclusion of nBSP in such cultures showed no effect on type I collagen telopeptide fragment release, hence overall bone resorption, whereas addition of dBSP abolished the PTH-induced bone resorption. The inhibition of bone resorption by dBSP was shown to be unique since in complementary experiments use of integrin receptor binding ligand, GRGDS peptide, offered only partial reduction on overall bone resorption. Quantitative RANKL analysis indicated that mechanistically the PTH-induced bone resorption was inhibited by dBSP via down-regulation of the osteoblastic RANKL production. This conclusion was supported by the RANKL analysis in cultured MC3T3-E1 osteoblast cells. Overall, these studies provided direct evidence for the involvement of covalently-bound phosphates on BSP in receptor mediated ‘outside-in’ signaling via transmembrane tyrosine phosphorylation with concurrent effects on down-stream gene expressions. The use of a live bone organ culture system augmented these results with further evidence that links the observed in vivo variable state of phosphorylation with bone remodeling.
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与骨吸收调节有关的破骨细胞功能抗原在生化上与玻染素受体有关。
DOI：
10.1083/jcb.109.4.1817
发表时间：
1989-10
期刊：
The Journal of cell biology
影响因子：
--
作者：
Davies J;Warwick J;Totty N;Philp R;Helfrich M;Horton M
通讯作者：
Horton M
影响因子：
4.2
作者：
Gericke, A;Qin, C;Boskey, AL
通讯作者：
Boskey, AL
影响因子：
4.1
作者：
Gordon, Jonathan A. R.;Tye, Coralee E.;Goldberg, Harvey A.
通讯作者：
Goldberg, Harvey A.
影响因子：
4.1
作者：
FRANZEN, A;HEINEGARD, D
通讯作者：
HEINEGARD, D
影响因子：
3.4
作者：
Decup, F;Six, N;Goldberg, M
通讯作者：
Goldberg, M