Structural and functional studies on the human hepatic interleukin-6 receptor. Molecular cloning and overexpression in HepG2 cells.
Structural and functional studies on the human hepatic interleukin-6 receptor. Molecular cloning and overexpression in HepG2 cells.
复制标题
人肝白细胞介素6受体的结构和功能研究。
作者：
H. Schooltink;T. Stoyan;D. Lenz;H. Schmitz;T. Hirano;T. Kishimoto;P. Heinrich;S. Rose
从地塞米松处理的人肝癌细胞(HepG2)的Poly(A)RNA文库中分离到编码人肝白细胞介素6受体(IL-6-R)的cDNA。我们发现肝脏IL-6-R与白细胞表达的IL-6-R相同。在兔体内制备了抗IL-6-R蛋白的多克隆抗血清。虽然整个IL-6-R蛋白被用于免疫，但只获得了针对IL-6-R胞浆区域的抗体。通过亲和交联和随后的免疫沉淀与抗配体和抗受体的抗体的亲和交联性和免疫沉淀法证明，所克隆的cDNA编码在HepG2细胞上具有功能的IL-6-R。当肝脏IL-6-R基因在HepG2细胞中过表达时，这些细胞对低浓度IL-6在诱导γ-纤维蛋白原mRNA方面变得更加敏感。
cDNAs coding for the human hepatic interleukin-6 receptor (IL-6-R) have been isolated from a library made from poly(A) RNA of dexamethasone-treated human hepatoma cells (HepG2). We found the hepatic IL-6-R to be identical to the one expressed by leucocytes. A polyclonal antiserum was raised in rabbits against the IL-6-R protein expressed in Escherichia coli. Although the entire IL-6-R protein was used for immunization, only antibodies to the cytoplasmic domain of the IL-6-R were obtained. It is demonstrated by affinity cross-linking and subsequent immunoprecipitation with antibodies against the ligand as well as against the receptor that the cloned cDNA codes for the functional IL-6-R on HepG2 cells. When the hepatic IL-6-R cDNA was overexpressed in HepG2 cells, these cells became more sensitive to low concentrations of IL-6 with respect to the induction of gamma-fibrinogen mRNA.