The P-selectin glycoprotein ligand functions as a common human leukocyte ligand for P- and E-selectins
The P-selectin glycoprotein ligand functions as a common human leukocyte ligand for P- and E-selectins
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P-选择素糖蛋白配体作为 P-和 E-选择素的常见人类白细胞配体
DOI：
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发表时间：
1995
影响因子：
4.8
通讯作者：
J. Geng
中科院分区：
文献类型：
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作者：
D. Asa;L. Raycroft;L. Ma;P. Aeed;PS Kaytes;AP Elhammer;J. Geng
P-和E-选择素属于Ca（2+）依赖性凝集素家族，并且作为髓系白细胞的受体起作用。我们已经描述了一组单克隆抗体，其识别来自人中性粒细胞和HL-60早幼粒细胞的唾液酸糖蛋白，并抑制这些细胞与P-选择素的粘附。在本研究中，我们发现E-选择素受体-球蛋白（E-selectin Rg）亲和层析可以以Ca（2+）依赖的方式从[3 H]葡糖胺标记的HL-60细胞中特异性地分离出一种糖蛋白。该蛋白在还原条件下具有约120 kDa的分子量，这似乎与先前表征的P-选择素的糖蛋白配体相同。该分子可以被E-和P-选择素柱交叉耗尽并与其交叉结合。通过P-和E-选择素亲和色谱法纯化的分子的去唾液酸化O-连接碳水化合物的色谱图是相同的。两者都有12.8、9.8、6.3、3.5和2.5个葡萄糖单位的五种结构。针对P-选择素唾液酸糖蛋白配体的PL 5单克隆抗体、E-选择素Rg和针对P-选择素糖蛋白配体-1（PSGL-1）的抗血清都在配体印迹和免疫印迹上识别纯化的P-选择素配体。此外，PL 5单克隆抗体阻断HL-60细胞和人中性粒细胞与E-选择素Rg的粘附。两者合计，我们的结果表明，P-和E-选择素配体在这项研究中定义的PSGL-1，并表明，这种分子是一个重要的白细胞配体的P-和E-选择素。
P- and E-selectins belong to a family of Ca(2+)-dependent lectins and function as receptors for myeloid leukocytes. We have described a panel of monoclonal antibodies which recognize a sialoglycoprotein from human neutrophils and HL-60 promyelocytic cells and inhibit adhesion of these cells to P-selectin. In this study, we show that the E-selectin receptor-globulin (E-selectin Rg) affinity chromatography can isolate specifically only one glycoprotein from [3H]glucosamine-labeled HL-60 cells in a Ca(2+)-dependent manner. This protein has a molecular mass of approximately 120 kDa under reducing conditions, which appears to be identical with the previously characterized glycoprotein ligand for P- selectin. The molecule can be cross-depleted by and cross-bound to the E- and P-selectin columns. The chromatographic profile of desialylated O-linked carbohydrates from molecules purified by P- and E-selectin affinity chromatography are identical. Both have five structures at 12.8, 9.8, 6.3, 3.5, and 2.5 glucose units. PL5 monoclonal antibody to the P-selectin sialoglycoprotein ligand, E-selectin Rg, and antiserum to P-selectin glycoprotein ligand-1 (PSGL-1) all recognize the purified P-selectin ligand on ligand blots and immunoblots. Furthermore, PL5 monoclonal antibody blocks adhesion of HL-60 cells and human neutrophils to E-selectin Rg. Taken together, our results demonstrate that the P- and E-selectin ligand defined in this study is PSGL-1 and suggest that this molecule is an important leukocyte ligand for both P- and E-selectins.