Binding of ADAM28 to P-selectin Glycoprotein Ligand-1 Enhances P-selectin-mediated Leukocyte Adhesion to Endothelial Cells*
Binding of ADAM28 to P-selectin Glycoprotein Ligand-1 Enhances P-selectin-mediated Leukocyte Adhesion to Endothelial Cells*
复制标题
DOI：
10.1074/jbc.m702414200
复制
发表时间：
2007-08
影响因子：
4.8
通讯作者：
M. Shimoda;G. Hashimoto;S. Mochizuki;E. Ikeda;N. Nagai;S. Ishida;Y. Okada
中科院分区：
文献类型：
--
作者：
M. Shimoda;G. Hashimoto;S. Mochizuki;E. Ikeda;N. Nagai;S. Ishida;Y. Okada
亚当斯（disintegrin and metalloproteinases）是一个新近发现的具有去整合素样结构域和金属蛋白酶结构域的多功能蛋白质基因家族。为了分析ADAM 28的生物学功能，我们利用酵母双杂交系统筛选了分泌型ADAM 28（ADAM 28 s）的结合分子，并鉴定了P-选择素糖蛋白配体-1（PSGL-1）。通过酵母双杂交试验、使用PSGL-1稳定转染子和表达天然PSGL-1的白细胞细胞系（HL-60细胞和Jurkat细胞）的结构域特异性重组ADAM 28物种的结合试验以及这些细胞中分子的共免疫定位和共免疫沉淀，测定ADAM 28 s的去整合素样结构域与PSGL-1的细胞外部分之间的结合。与重组ADAM 28共同孵育HL-60细胞可增强与P-选择素包被的威尔斯孔和表达P-选择素的内皮细胞的结合。此外，在内毒素诱导的炎症小鼠模型中，静脉注射ADAM 28处理的HL-60细胞增加了它们在肺微循环和肺泡腔中的积累。这些数据表明了一种新的功能，即在炎症条件下，ADAM 28 s通过与白细胞上的PSGL-1相互作用促进PSGL-1/P-选择素介导的白细胞滚动粘附至内皮细胞并随后浸润至组织间隙。
ADAMs (a disintegrin and metalloproteinases) are a recently discovered gene family of multifunctional proteins with the disintegrin-like and metalloproteinase domains. To analyze the biological functions of ADAM28, we screened binding molecules to secreted-type ADAM28 (ADAM28s) by the yeast two-hybrid system and identified P-selectin glycoprotein ligand-1 (PSGL-1). Binding between the disintegrin-like domain of ADAM28s and the extracellular portion of PSGL-1 was determined by yeast two-hybrid assays, binding assays of the domain-specific recombinant ADAM28s species using PSGL-1 stable transfectants and leukocyte cell lines expressing native PSGL-1 (HL-60 cells and Jurkat cells), and co-immunolocalization and co-immunoprecipitation of the molecules in these cells. Incubation of HL-60 cells with recombinant ADAM28s enhanced the binding to P-selectin-coated wells and P-selectin-expressing endothelial cells. In addition, intravenous injection of ADAM28s-treated HL-60 cells increased their accumulation in the pulmonary microcirculation and alveolar spaces in a mouse model of endotoxin-induced inflammation. These data suggest a novel function that ADAM28s promotes PSGL-1/P-selectin-mediated leukocyte rolling adhesion to endothelial cells and subsequent infiltration into tissue spaces through interaction with PSGL-1 on leukocytes under inflammatory conditions.