Isolation and characterization of a Clostridium botulinum C2 toxin-resistant cell line: evidence for possible involvement of the cellular C2II receptor in growth regulation
Isolation and characterization of a Clostridium botulinum C2 toxin-resistant cell line: evidence for possible involvement of the cellular C2II receptor in growth regulation
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肉毒梭菌 C2 毒素抗性细胞系的分离和表征：细胞 C2II 受体可能参与生长调节的证据
DOI：
10.1128/iai.63.6.2334-2340.1995
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发表时间：
1995
影响因子：
3.1
通讯作者：
K. Aktories
中科院分区：
文献类型：
--
作者：
G. Fritz;P. Schroeder;K. Aktories
肉毒杆菌C2毒素由结合组分C2II和酶组分C2I组成，通过g -肌动蛋白的选择性adp核糖基化作用于真核细胞。为了获得C2耐药细胞，我们用n-亚硝基-n -甲基脲诱变CHO-K1细胞，并选择C2耐药细胞。在C2I浓度为50 ng / ml和C2II浓度为100 ng / ml的条件下，以30 × 10（-6）的频率获得存活的细胞。当C2I浓度为10 ng / ml， C2II浓度为20 ng / ml时，CHO野生型细胞的集落形成能力降低到50%。相比之下，分离的CHO突变细胞的集落形成能力不受C2I浓度为200 ng / ml和C2II浓度为400 ng / ml的影响。毒素诱导的g -肌动蛋白adp核糖基化在突变细胞的裂解物中没有受损。C2抗毒素表型对产气荚膜梭菌、新梭菌、艰难梭菌和肉毒梭菌（C3）的细胞毒素和细胞松弛素d的细胞围住活性保持敏感，在抗性CHO细胞中，组分C2II的结合受损，提示毒素细胞表面受体发生突变。血清因子保护野生型细胞免受C2毒素的细胞毒性作用。此外，c2抗性表型与血清依赖性增加相关。这些数据表明，肉毒杆菌C2毒素的作用是通过其结合和摄取细胞表面受体介导的，该受体可能参与生长调节。
Clostridium botulinum C2 toxin, which consists of the binding component C2II and the enzyme component C2I, acts on eukaryotic cells by selective ADP-ribosylation of G-actin. To obtain C2 toxin-resistant cells, we mutagenized CHO-K1 cells with N-nitroso-N-methylurea and selected for C2 resistance. Cells which survived the selection procedure with 50 ng of C2I and 100 ng of C2II per ml were obtained with a frequency of 30 x 10(-6). The colony-forming ability of CHO wild-type cells was reduced to 50% with 10 ng of C2I and 20 ng of C2II per ml. In contrast, the colony-forming ability of the isolated CHO mutant cells was not influenced by up to 200 ng of C2I and 400 ng of C2II per ml. Toxin-induced ADP-ribosylation of G-actin was not impaired in lysates of mutant cells. The C2 toxin-resistant phenotype remained sensitive to the cell-rounding activities of cytotoxins from C. perfringens (iota-toxin), C. novyi, C. difficile, and C. botulinum (C3) and to cytochalasin D. Binding of component C2II was impaired in resistant CHO cells, suggesting mutation of the toxin cell surface receptor. Serum factors protected wild-type cells against the cytotoxic effect of C2 toxin. Furthermore, the C2-resistant phenotype correlated with an increased serum dependency. The data suggest that the action of C. botulinum C2 toxin is mediated by its binding and uptake via a cell surface receptor which might be involved in growth regulation.
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DOI：
10.1016/0041-0101(86)90101-7
发表时间：
1986
期刊：
Toxicon : official journal of the International Society on Toxinology
影响因子：
--
作者：
Stiles,BG;Wilkins,TD
通讯作者：
Wilkins,TD
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DOI：
10.1073/pnas.87.18.7250
发表时间：
1990
影响因子：
11.1
作者：
Naglich,JG;Eidels,L
通讯作者：
Eidels,L
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DOI：
--
发表时间：
1989
期刊：
The Journal of pharmacology and experimental therapeutics
影响因子：
--
作者：
Simpson,LI
通讯作者：
Simpson,LI
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DOI：
--
发表时间：
1984
期刊：
The Journal of pharmacology and experimental therapeutics
影响因子：
--
作者：
Simpson,LL
通讯作者：
Simpson,LL