Scheving L A, Shiurba R A, Nguyen T D, Gray G M
Department of Medicine (Gastroenterology Division), Stanford University School of Medicine, California 94305.
J Biol Chem. 1989 Jan 25;264(3):1735-41.
Interaction of epidermal growth factor (EGF) with its specific receptor (EGFR) was explored in the intact rat small intestine and in highly purified isolated enterocyte membrane preparations. Despite the fact that the EGF ligand is known to be present at physiological concentrations within the intestinal cavity, no significant binding of the ligand to the brush border surface was observed. Instead, binding of EGF to the EGFR was confined to other membrane populations, and correlation of ligand interaction with the laterobasal membranes (LBM) was nearly perfect (p less than 0.001) across a special equilibrium gradient enriched in brush border and LBM but devoid of intracellular membranes. Specific binding to another minor population of intracellular membranes that migrated to a position less dense than typical endoplasmic reticulum-Golgi vesicles on equilibrium gradients was also observed. Immunocytochemical exposure of intestine to EGFR antibody confirmed the localization of the EGFR to LBM and intracellular membranes. As estimated from the intensity of the staining, there may be immunologically active but nonbinding receptor species in the intracellular membrane compartment. Thus, despite the secretion of EGF into the intestinal lumen, the growth and maturational effects of EGF probably result from a specific interaction between EGF and EGFR solely at the laterobasal surface of the enterocyte. The functional role of the intracellular membrane species of EGFR, which remains to be established, may involve a source of inactive receptor that can be rapidly recruited and transferred to the LBM surface under changing environmental conditions.
在完整的大鼠小肠和高度纯化的分离肠细胞膜制剂中,研究了表皮生长因子(EGF)与其特异性受体(EGFR)的相互作用。尽管已知EGF配体在肠腔内以生理浓度存在,但未观察到该配体与刷状缘表面有明显结合。相反,EGF与EGFR的结合局限于其他膜群体,并且在富含刷状缘和侧基底膜(LBM)但无细胞内膜的特殊平衡梯度中,配体与LBM的相互作用相关性近乎完美(p小于0.001)。还观察到EGF与另一小部分细胞内膜有特异性结合,这些细胞内膜在平衡梯度上迁移到比典型内质网-高尔基体囊泡密度更低的位置。用EGFR抗体对小肠进行免疫细胞化学检测,证实了EGFR定位于LBM和细胞内膜。根据染色强度估计,细胞内膜区室中可能存在具有免疫活性但不结合的受体种类。因此,尽管EGF分泌到肠腔中,但EGF的生长和成熟作用可能仅源于EGF与EGFR在肠细胞侧基底表面的特异性相互作用。EGFR细胞内膜种类的功能作用尚待确定,可能涉及一种无活性受体的来源,在不断变化的环境条件下,它可以迅速被募集并转移到LBM表面。