Effect of surface-adsorbed proteins and phosphorylation inhibitor AG18 on intracellular protein expression in adherent macrophages.
Effect of surface-adsorbed proteins and phosphorylation inhibitor AG18 on intracellular protein expression in adherent macrophages.
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表面吸附蛋白和磷酸化抑制剂 AG18 对贴壁巨噬细胞胞内蛋白表达的影响。
DOI：
10.1016/j.biomaterials.2006.02.028
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发表时间：
2006
期刊：
影响因子：
14
通讯作者：
Kao,WeiyuanJohn
中科院分区：
文献类型：
--
作者：
Zuckerman,SeanT;Kao,WeiyuanJohn
巨噬细胞被认为在宿主对植入生物材料的炎症反应中起重要作用。然而，巨噬细胞粘附于蛋白质吸附底物以及随后的激活和炎症的机制尚不清楚。在此之前，我们使用SDS-PAGE和密度测定法鉴定了各种表面吸附蛋白和磷酸化抑制剂AG18浓度增加对贴壁人单核细胞系U937细胞内蛋白表达水平的影响。蛋白质配体和AG18浓度上调或下调一系列蛋白的表达，范围从~ 200到~ 23kDa。本研究采用高效液相色谱-串联质谱（HPLC - coupled tandem mass spectrometry, LC/MS）对这些条带中的蛋白质进行了鉴定。我们假设巨噬细胞粘附和活化的关键蛋白可以通过观察不同表面吸附配体和AG18浓度引起的蛋白表达来确定。在~ 52、~ 42和~ 23kDa时，AG18浓度下调或上调可调节贴壁U937在pbs吸附的TCPS上的蛋白表达。AG18浓度对白蛋白（Alb）吸附表面的细胞没有影响，但在40和80μm AG18下，可调节纤维连接蛋白（FN）吸附的TCPS上粘附U937的不同蛋白表达。Alb和FN作为表面吸附配体调节贴壁细胞中不同的蛋白质。根据LC/MS的数据，表面相关配体和AG18浓度的增加都可以调节细胞内信号传导的变化。
Macrophages are believed to play an important role in the host inflammatory response to implanted biomaterials. However, the mechanism of macrophage adhesion to protein-adsorbed substrates and the subsequent activation and inflammation is unresolved. Previously the effect of various surface-adsorbed proteins and increasing concentrations of phosphorylation inhibitor AG18 on intracellular protein expression levels in adherent human monocytic cell line U937 was identified using SDS-PAGE and densitometry. The protein ligands and AG18 concentrations up or down regulated the expression of a set of proteins ranging from ∼200 to ∼23kDa. In the present work, HPLC coupled tandem mass spectroscopy (LC/MS) was used to identify proteins in these bands. We hypothesized that key proteins in macrophage adhesion and activation could be identified by observing protein expression resulting from various surface-adsorbed ligands and AG18 concentrations. Increasing concentrations of AG18 down or up regulate protein expression in adherent U937 on PBS-adsorbed TCPS at ∼52, ∼42 and ∼23kDa. AG18 concentrations had no effect on cells on albumin (Alb)-adsorbed surfaces but regulated different protein expression in adherent U937 on fibronectin (FN)-adsorbed TCPS at 40 and 80μm AG18. Both Alb and FN regulate distinct sets of proteins in adherent cells as surface-adsorbed ligands. Based on the data from LC/MS, both surface associated ligand and increasing concentrations of AG18 modulate shifts in intracellular signaling.
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蛋白质组学、主成分分析和转录组学的结合是鉴定 U937 细胞系中巨噬细胞成熟生物标志物的强大工具
影响因子：
3.4
作者：
K. Verhoeckx;S. Bijlsma;E. M. de Groene;R. Witkamp;J. van der Greef;R. Rodenburg
通讯作者：
R. Rodenburg
用于组织分析的新型蛋白质组学方法
DOI：
--
发表时间：
2004
期刊：
Espert Review of Proteomics
影响因子：
--
作者：
M. Tangrea;Benjamin S. Wallis;J. Gillespie;G. Gannot;M. Emmert;R. Chuaqui
通讯作者：
R. Chuaqui
影响因子：
13.8
作者：
DSOUZA, SE;GINSBERG, MH;PLOW, EF
通讯作者：
PLOW, EF
整合素 α5β1 调节 αvβ3 介导的细胞外信号调节激酶激活1
DOI：
--
发表时间：
2005
期刊：
影响因子：
--
作者：
D. Ly;S. Corbett
通讯作者：
S. Corbett