PURIFICATION AND SOME PROPERTIES OF A VERO TOXIN FROM A HUMAN STRAIN OF ESCHERICHIA-COLI THAT IS IMMUNOLOGICALLY RELATED TO SHIGA-LIKE TOXIN-II (VT2)
PURIFICATION AND SOME PROPERTIES OF A VERO TOXIN FROM A HUMAN STRAIN OF ESCHERICHIA-COLI THAT IS IMMUNOLOGICALLY RELATED TO SHIGA-LIKE TOXIN-II (VT2)
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DOI：
10.1016/0882-4010(89)90014-4
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发表时间：
1989-02-01
影响因子：
3.8
通讯作者：
TAKEDA, Y
中科院分区：
文献类型：
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作者：
OKU, Y;YUTSUDO, T;TAKEDA, Y
从溶血性尿毒症综合征患者分离的大肠杆菌菌株中纯化出一种与志贺样毒素 II (SLT-II)（或 VT2）免疫学相关的 Vero 细胞细胞毒素（Vero 毒素）。该毒素对 Vero 细胞有活性，但对 HeLa 细胞活性低得多，这一特性与最近发现的大肠杆菌菌株中引起猪水肿病的 SLT-II 变体的特性相似。因此本报告纯化的毒素暂命名为志贺样毒素II变体（Vero毒素2变体）。纯化程序包括硫酸铵分级、DEAE-Sepharose CL-6B 柱色谱、色谱聚焦柱色谱以及在 TSK-gel G-2000SW 柱和 TSK-gel DEAE-5PW 柱上重复高效液相色谱 (HPLC)。从451份培养物上清液中获得约90μg纯化的毒素，产率约16％。纯化的毒素由分子大小与 SLT-II (VT2) 相似的 A 和 B 亚基组成。纯化毒素的等电点为6.1，与SLT-II (VT2)不同(pl = 4.1)。在 Ouchterlony 双凝胶扩散试验中，纯化的毒素和 STL-II (VT2) 形成沉淀素线，并分别针对抗纯化的毒素和抗 SLT-II (抗 VT2) 形成刺突。纯化的毒素对Vero细胞具有细胞毒性，约6pg的毒素可杀死50%的Vero细胞，腹腔注射时对小鼠表现出致死毒性，LD50为每只小鼠约2.7ng。
A cytotoxin to Vero cells (Vero toxin), which was immunologically related to Shiga-like toxin II (SLT-II) (or VT2), was purified from a strain of Escherichia coli isolated from a patient with hemolytic uremic syndrome. The toxin was active on Vero cells but much less active on HeLa cells, a property similar to that of the recently identified SLT-II variant from E. coli strains that caused edema disease of swine. Thus the toxin purified in this report was tentatively named Shiga-like toxin II variant (Vero toxin 2 variant). The purification procedures consisted of ammonium sulfate fractionation, DEAE-Sepharose CL-6B column chromatography, chromatofocusing column chromatography, and repeated high performance liquid chromatography (HPLC) on TSK-gel G-2000SW column and on TSK-gel DEAE-5PW columns. About 90 .mu.g of purified toxin was obtained from 451 of the culture supernatant with a yield of about 16%. The purified toxin consisted of A and B subunits of molecular sizes similar to those of SLT-II (VT2). The isoelectric point of the purified toxin was 6.1, which was different from that of SLT-II (VT2) (pl = 4.1). In an Ouchterlony double gel diffusion test, purified toxin and STL-II (VT2) formed precipitin lines with spur formation against anti-purified toxin and anti-SLT-II (anti-VT2), respectively. The purified toxin was cytotoxic to Vero cells, about 6 pg of the toxin killing 50% of the Vero cells, and showed lethal toxicity to mice when injected intraperitoneally, the LD50 being about 2.7 ng per mouse.