Chen Y H, Böck G, Vornhagen R, Steindl F, Katinger H, Dierich M P
Ludwig-Boltzmann-Institute of AIDS Research, Innsbruck, Austria.
Mol Immunol. 1993 Sep;30(13):1159-63. doi: 10.1016/0161-5890(93)90134-w.
Based on our findings that HIV-1 gp41 independently of CD4, can bind to the helper T-cell line H9, we characterized putative binding of HIV-1 gp41 to B-cell lines, Raji, Bjab and Ramos. Using fluorescence-activated cell sorter (FACS) we examined the binding of soluble gp41 (sgp41; Env amino acid 539-684) to these B-cell lines. Using sgp41 attached to sepharose beads Raji cell lysates were absorbed. The sgp41-eluate of Raji cell lysates could inhibit the sgp41-binding to Raji cells. By SDS-PAGE of sgp41-eluate of Raji cell lysates four strong protein band, 37, 45, 49 and 62 kD, and a weak band of 92 kD were stained with Coomassie blue. By Western blot (ligand blot) analysis using sgp41 four protein bands, 37, 45, 49 and 62 kD, were observed in sgp41-eluate of Raji cell lysates. To test the individual proteins the five proteins were isolated from the sgp41-eluate of Raji cell lysates. Three proteins, 45, 49 and 62 kD, each could partially inhibit the sgp41-binding to Raji cells. The results suggest that these three proteins in Raji cell lysates are possible candidates for the putative gp41 receptor(s).
基于我们的研究发现,即HIV-1 gp41可独立于CD4与辅助性T细胞系H9结合,我们对HIV-1 gp41与B细胞系Raji、Bjab和Ramos的假定结合进行了表征。我们使用荧光激活细胞分选仪(FACS)检测了可溶性gp41(sgp41;Env氨基酸539 - 684)与这些B细胞系的结合。使用连接到琼脂糖珠上的sgp41对Raji细胞裂解物进行吸附。Raji细胞裂解物的sgp41洗脱液可抑制sgp41与Raji细胞的结合。通过对Raji细胞裂解物的sgp41洗脱液进行SDS - PAGE,考马斯亮蓝染色显示出四条强蛋白带,分别为37、45、49和62 kD,以及一条92 kD的弱带。通过使用sgp41进行蛋白质印迹(配体印迹)分析,在Raji细胞裂解物的sgp41洗脱液中观察到四条蛋白带,分别为37、45、49和62 kD。为了测试单个蛋白质,从Raji细胞裂解物的sgp41洗脱液中分离出这五种蛋白质。三种蛋白质,45、49和62 kD,每种都可部分抑制sgp41与Raji细胞的结合。结果表明,Raji细胞裂解物中的这三种蛋白质可能是假定的gp41受体的候选者。