MURINE ENDOTHELIAL LEUKOCYTE-ADHESION MOLECULE-1 IS A CLOSE STRUCTURAL AND FUNCTIONAL HOMOLOG OF THE HUMAN PROTEIN
MURINE ENDOTHELIAL LEUKOCYTE-ADHESION MOLECULE-1 IS A CLOSE STRUCTURAL AND FUNCTIONAL HOMOLOG OF THE HUMAN PROTEIN
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DOI：
10.1111/j.1432-1033.1992.tb16940.x
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发表时间：
1992-06-01
期刊：
影响因子：
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通讯作者：
DELAMARTER, JF
中科院分区：
文献类型：
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作者：
BECKERANDRE, M;VANHUIJSDUIJNEN, RH;DELAMARTER, JF
人内皮细胞白细胞粘附分子1（ELAM-1）是一种仅表达于血管内皮细胞表面的糖蛋白，介导中性粒细胞、记忆性T细胞和单核细胞的粘附。ELAM-1，也称为E-选择素或白细胞内皮细胞粘附分子2，是凝集素/表皮生长因子/补体调节蛋白样细胞粘附分子家族的成员，其包括称为选择素的结构相关分子。它们都参与细胞/细胞粘附，在白细胞运输中发挥作用，目前仅部分确定。我们在这里报告的人ELAM-1的鼠等效物的分离和表征。小鼠ELAM-1由一个单拷贝基因编码，跨越约13 kb，其在结构上被组织成14个外显子和13个内含子;与其人类对应物非常相似。外显子/内含子结构与所编码蛋白质的结构域完全平行。从白细胞介素-1-（IL-1）处理的小鼠的心脏组织中克隆了鼠ELAM-1特异性cDNA。其核苷酸序列与人ELAM-1 cDNA的相似性为70%。在Cos细胞中瞬时表达，编码的蛋白质促进重组细胞与人多态性核细胞以及表达S-Lewis（-x）糖部分的HL 60细胞之间的粘附。北方印迹研究揭示了迄今为止小鼠ELAM-1基因在IL-1处理的小鼠的心脏组织中的最高表达和仅在肺组织中的低表达。在启动子内，大多数最近鉴定的调控元件是保守的。一个例外是核因子（NF）κ-B盒序列，其在鼠ELAM-1启动子中不对应于共有NF-κ-B序列（Lenardo和巴尔的摩，1989）。带移分析显示与NF-κ B样蛋白没有结合。然而，与人ELAM-1启动子相比，鼠ELAM-1启动子与氯霉素乙酰转移酶报告基因的融合赋予了精氨酸诱导的转录，尽管水平较低。我们的研究结果表明，存在的人基因的小鼠同源物，并证明从两个物种的同源蛋白质之间的粘附功能等效。此外，我们提供了第一个证据的效用的小鼠模型在解决生物学问题的作用，ELAM-1在炎症中发挥。
Human endothelial leukocyte-adhesion molecule 1 (ELAM-1), a cell-surface glycoprotein expressed solely on cytokine-activated endothelial cells, mediates the adhesion of blood neutrophils, memory T-cells and some monocytes. ELAM-1, also known as E-selectin or leukocyte endothelial-cell-adhesion molecule 2, is a member of the lectin/epidermal-growth-factor/complement-regulatory-protein-like cell-adhesion molecule family, which includes structurally related molecules referred to as selectins. They are all involved in cell/cell adhesion, playing roles in leukocyte trafficking which are currently only partially defined. We report here the isolation and characterization of the murine equivalent of human ELAM-1. Murine ELAM-1 is encoded by a single-copy gene, spanning about 13 kb, which is structurally organized into 14 exons and 13 introns; very similar to that of its human counterpart. The exon/intron architecture exactly parallels the domain structure of the encoded protein. A murine ELAM-1-specific cDNA was cloned from heart tissue of an interleukin-1-(IL-1)-treated mouse. Its nucleotide sequence shows an overall similarity of 70% to human ELAM-1 cDNA. Transiently expressed in Cos cells, the encoded protein promotes the adhesion between recombinant cells and both human polymorphic nuclear cells, as well as HL60 cells expressing the S-Lewis(-x) sugar moiety. Northern blot studies revealed by far the highest expression of the murine ELAM-1 gene in heart tissue and only low expression in lung tissue of IL-1-treated mice. Within the promoter, most of the recently identified regulatory elements are conserved. An exception is the nuclear factor (NF)kappa-B box sequence, which, in the murine ELAM-1 promoter, does not correspond to the consensus NF-kappa-B sequence (Lenardo and Baltimore, 1989). Band-shift analyses show no binding to NF-kappa-B-like proteins. However, fusion of the murine ELAM-1 promoter to a chloramphenicol acetyltransferase reporter confers cytokine-inducible transcription, although at a lower level, when compared to the human ELAM-1 promoter. Our results demonstrate the existence of a murine homologue of the human gene and demonstrate for adhesion functional equivalence between the homologous proteins from the two species. In addition, we provide the first evidence of the utility of the murine model in addressing biological questions about the role which ELAM-1 plays in inflammation.