DIPHTHERIA-TOXIN RECEPTOR-BINDING DOMAIN SUBSTITUTION WITH INTERLEUKIN-6 - GENETIC CONSTRUCTION AND INTERLEUKIN-6 RECEPTOR-SPECIFIC ACTION OF A DIPHTHERIA TOXIN-RELATED INTERLEUKIN-6 FUSION PROTEIN

被引:33
作者
JEAN, LFL
MURPHY, JR
机构
[1] BOSTON UNIV HOSP,EVANS DEPT CLIN RES,88 E NEWTOWN ST,BOSTON,MA 02218
[2] BOSTON UNIV,SCH MED,DEPT MICROBIOL,BOSTON,MA 02118
[3] BOSTON UNIV HOSP,DEPT MED,BOSTON,MA 02218
来源
PROTEIN ENGINEERING | 1991年 / 4卷 / 08期
关键词
DIPHTHERIA TOXIN; EUKARYOTIC CELLS; INTERLEUKIN-6;
D O I
10.1093/protein/4.8.989
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We have genetically replaced that portion of the diphtheria toxin structural gene which encodes the native receptor-binding domain with a synthetic gene encoding the cytokine interleukin 6 (IL-6/IFN-beta-2/BSF-2). The resulting gene fusion encodes the chimeric toxin DAB389-IL-6. Following expression and purification, we demonstrate that DAB389-IL-6 is selectively cytotoxic for eukaryotic cells bearing the interleukin 6 receptor. In addition, the cytotoxic action of DAB389-IL-6 is shown to require binding to the IL-6 receptor, internalization by receptor-mediated endocytosis and passage through an acidic compartment. Following the delivery of the catalytically active fragment A to the cytosol of target cells, cellular protein synthesis is inhibited by the ADP-ribosylation of elongation factor 2. While eukaryotic cells which are devoid of the IL-6 receptor are uniformly resistant to the action of this fusion toxin, the data presented suggest that a minimal number of IL-6 receptors may be necessary to mediate the internalization of sufficient levels of DAB389-IL-6 to result in the intoxication of target cells.
引用
收藏
页码:989 / 994
页数:6
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