Fecondo J V, Kent S B, Boyd A W
Lions Clinical Cancer Research Laboratory, Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia.
Proc Natl Acad Sci U S A. 1991 Apr 1;88(7):2879-82. doi: 10.1073/pnas.88.7.2879.
We have used a combination of hydropathy analysis of the intercellular adhesion molecule 1 (ICAM-1) sequence and dot-matrix comparison of the sequence with the homologous, but functionally distinct, protein myelin-associated glycoprotein to identify a putative functional binding region. One polar, and presumably surface-exposed, region of ICAM-1 showed no significant identity with myelin-associated glycoprotein. A synthetic peptide analog based on the sequence of this region (JF9) mimicked the inhibitory effects of the anti-ICAM-1 monoclonal antibody WEHI-CAM-1. These included inhibition of ICAM-1-dependent homotypic aggregation of Raji Burkitt lymphoma and phorbol-ester treated U937 cells at concentrations as low as 80 micrograms/ml (24 microM). In addition, at a concentration of 100 micrograms/ml, the peptide analog effectively inhibited cytotoxic cell activity, an ICAM-1-dependent effector function of the immune response. This simple method of sequence analysis may have general applicability to the identification of functional domains in homologous, but functionally distinct, proteins such as the translated products of gene families.
我们通过对细胞间黏附分子1(ICAM-1)序列进行亲水性分析,并将该序列与同源但功能不同的蛋白质髓鞘相关糖蛋白进行点阵比较,以确定一个假定的功能结合区域。ICAM-1的一个极性且可能暴露于表面的区域与髓鞘相关糖蛋白没有显著的一致性。基于该区域序列(JF9)的合成肽类似物模拟了抗ICAM-1单克隆抗体WEHI-CAM-1的抑制作用。这些作用包括在低至80微克/毫升(24微摩尔)的浓度下抑制Raji伯基特淋巴瘤细胞和佛波酯处理的U937细胞的ICAM-1依赖性同型聚集。此外,在100微克/毫升的浓度下,该肽类似物有效抑制细胞毒性细胞活性,这是免疫反应中一种ICAM-1依赖性效应功能。这种简单的序列分析方法可能普遍适用于鉴定同源但功能不同的蛋白质(如基因家族的翻译产物)中的功能结构域。