Direct proteomic mapping of the lung microvascular endothelial cell surface in vivo and in cell culture
Direct proteomic mapping of the lung microvascular endothelial cell surface in vivo and in cell culture
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DOI：
10.1038/nbt993
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发表时间：
2004-08-01
影响因子：
46.9
通讯作者：
Schnitzer, JE
中科院分区：
文献类型：
--
作者：
Durr, E;Yu, JY;Schnitzer, JE
内皮细胞在体外和体内可以发挥不同的功能;然而，体内微环境调节的程度在分子水平上仍然未知，主要是因为分析的局限性。我们使用多维蛋白质鉴定技术（MudPIT），以确定450蛋白质（三个或更多个光谱）从大鼠肺和培养的大鼠肺微血管内皮细胞分离的管腔内皮细胞质膜。41%的体内表达蛋白在体外检测不到。测量再现性的统计分析表明，使用当前的离子阱设备，要实现大于或等于95%的分析完整性置信度，必须进行7到10次MudPIT测量。大规模的映射血管内皮细胞表面的蛋白质组在体内，如这里所示，是可取的，因为不同的蛋白质的表达显然是由组织微环境，还不能复制在标准的细胞培养。
Endothelial cells can function differently in vitro and in vivo; however, the degree of microenvironmental modulation in vivo remains unknown at the molecular level largely because of analytical limitations. We use multidimensional protein identification technology (MudPIT) to identify 450 proteins (with three or more spectra) in luminal endothelial cell plasma membranes isolated from rat lungs and from cultured rat lung microvascular endothelial cells. Forty-one percent of proteins expressed in vivo are not detected in vitro. Statistical analysis measuring reproducibility reveals that seven to ten MudPIT measurements are necessary to achieve greater than or equal to95% confidence of analytical completeness with current ion trap equipment. Large-scale mapping of the proteome of vascular endothelial cell surface in vivo, as demonstrated here, is advisable because distinct protein expression is apparently regulated by the tissue microenvironment that cannot yet be duplicated in standard cell culture.