DIFFERENTIAL EXPRESSION OF CELL-SURFACE GLYCOPROTEINS ON VARIOUS ORGAN-DERIVED MICROVASCULAR ENDOTHELIA AND ENDOTHELIAL-CELL CULTURES
DIFFERENTIAL EXPRESSION OF CELL-SURFACE GLYCOPROTEINS ON VARIOUS ORGAN-DERIVED MICROVASCULAR ENDOTHELIA AND ENDOTHELIAL-CELL CULTURES
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DOI：
10.1002/jcp.1041360303
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发表时间：
1988-09-01
影响因子：
5.6
通讯作者：
NICOLSON, GL
中科院分区：
文献类型：
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作者：
BELLONI, PN;NICOLSON, GL
对源自不同小鼠器官的微血管内皮管腔表面表达的糖蛋白进行分析，并与培养的血管内皮细胞表达的糖蛋白进行比较。通过心内灌注乳过氧化物酶/Na125I 对细胞表面血管蛋白进行原位放射性标记。放射自显影证实放射性标记仅限于大多数组织的血管腔。对照含有 125I 标记的血清蛋白，以识别吸附的血清成分。使用从不同器官分离的 125I 标记微血管的洗涤剂提取物，通过西方酶联凝集素分析来分析糖蛋白。用一组凝集素-过氧化物酶缀合物探测蛋白质印迹，以确定蛋白质糖基化的差异。还通过放射自显影术筛选相同的转移物中暴露的 125I 标记的细胞表面蛋白。这种双重分析检测到每个器官独特的糖蛋白模式。至少七种主要蛋白质（Mr.大约180K、130K、95K、80K、75K、60K、12K）是源自每个器官的微血管所共有的；然而，某些糖蛋白似乎在特定器官中存在差异表达。例如，.apprx 先生。在脑微血管中检测到 135 K WGA 结合糖蛋白，而 Mr.apprx 的另一种 WGA 结合糖蛋白。仅在肾脏中检测到 40 K。在肺微血管中，.apprx 先生。 140 K WGA 结合糖蛋白和 Mr.apprx。 55 K RCA-I 结合半乳蛋白优先暴露，肝脏微血管显示 Mr.apprx。 220 K 蛋白质和 Mr.apprx。 35 K PNA 结合半乳蛋白。原位标记微血管的细胞表面碘化蛋白质谱与培养的牛主动脉内皮细胞和从不同器官分离的几种短期内皮细胞培养物的谱相似。这项研究的结果表明，器官相关内皮细胞表达每个器官特有的糖蛋白指纹。
Glycoproteins expressed on the luminal surfaces of microvascular endothelium derived from various murine organs were analyzed and compared with those expressed by cultured vascular endothelial cells. Cell-surface vascular proteins were radiolabeled in situ via intracardiac perfusion with lactoperoxidase/Na125I. Autoradiography confirmed that the radiolabel was restricted to the vessel lumen in most tissues. Controls contained 125I-labeled serum proteins to identify adsorbed serum components. Glycoproteins were analyzed by western enzyme-linked lectin analysis using detergent extracts of 125I-labeled microvessels isolated from different organs. The western transfers were probed with a panel of lectin-peroxidase conjugates to determine differences in protein glycosylation. The same transfers were also screened for exposed 125I-labeled cell-surface proteins by autoradiography. This dual analysis detected glycoprotein patterns unique for each organ. At least seven major proteins (Mr .apprx. 180 K, 130 K, 95 K, 80 K, 75 K, 60 K, 12 K) were common to microvessels derived from each organ; however, certain glycoproteins appeared to be expressed differentially in particular organs. For example, a Mr .apprx. 135 K WGA-binding glycoprotein was detected in brain microvessels, whereas another WGA-binding glycoprotein of Mr .apprx. 40 K was detected only in kidney. In lung microvessels, a Mr .apprx. 140 K WGA binding glycoprotein and a Mr .apprx. 55 K RCA-I-binding galactoprotein were exposed preferentially, and liver microvessels displayed Mr .apprx. 220 K protein and a Mr .apprx. 35 K PNA-binding galactoprotein. The cell-surface-iodinated protein profiles from in situ labeled microvessels were similar to profiles derived from cultured bovine aortic endothelial cells and several short-term endothelial cells cultures isolated from different organs. The results from this study suggest that organ-associated endothelia express glycoprotein fingerprints unique to each organ.