Structural analysis and identification of biological activity of alpha-toxin amino-terminal PLC from Clostridium welchii type A

doi:10.3969/j.issn.1004-1524.2017.02.06

›› 2017, Vol. 29 ›› Issue (2): 213-219.DOI: 10.3969/j.issn.1004-1524.2017.02.06

• Animal Science • Previous Articles Next Articles

Structural analysis and identification of biological activity of alpha-toxin amino-terminal PLC from Clostridium welchii type A

XU Chongli¹, XU Chongbo^{2, *}, GONG Yuchen³

1. College of Biology and Food Engineering, Jilin Institute of Chemical Technology, Jilin 132022, China;
2. North Guangdong Collaborative Innovation and Development Center for Swine Farming and Disease Control, Yingdong College of Life Sciences, Shaoguan University, Shaoguan 512005, China;
3. College of Animal Science, Jilin Agricultural University, Changchun 130118, China

Received:2016-09-26 Online:2017-02-15 Published:2017-03-06

Abstract

Abstract: Amino-terminal PLC1-250 gene of Clostridium welchii α-toxin was amplified by PCR. The recombinant strain BL21 (DE3)(pN-PLC1-250) containing PLC1-250 gene was constructed. It was shown that the recombinant plasmid pN-PLC1-250 contained PLC1-250 gene with correct sequence and ORF by identification of endonuclease-digesting and sequence analysis. SDS-PAGE analysis showed that the expression level of PLC1-250 proteins were about 18.76% of total cellular proteins. The secondary structure and three-dimensional structure of PLC1-250 proteins were predicted by SOPMA method on EXPASY website. The results showed that the secondary structure of PLC1-250 protein was composed of alpha helices and random coils. The three-dimensional structure of PLC1-250 protein was similar with amino-terminal domain of α-toxin protein. The study of biological activity laid foundation for further research of the molecular mechanism of α-toxin and the relation of its molecular structure and biological function.

Key words: Clostridium welchii type A, alpha-toxin PLC-gene, biological activity, circular dichroism spectra

CLC Number:

S852
Q78

XU Chongli, XU Chongbo, GONG Yuchen. Structural analysis and identification of biological activity of alpha-toxin amino-terminal PLC from Clostridium welchii type A[J]. , 2017, 29(2): 213-219.

References

[1] GOHARI I M, PARREIRA V R, NOWELL V J, et al. A novel pore-forming toxin in type A Clostridium perfringens is associated with both fatal canine hemorrhagic gastroenteritis and fatal foal necrotizing enterocolitis[J]. PLoS One, 2015, 10(4): e0122684.
[2] MONMA C, HATAKEYAMA K, OBATA H, et al. Four foodborne disease outbreaks caused by a new type of enterotoxin-producing Clostridium perfringens[J]. Journal of Clinical Microbiology, 2015, 53(3):859-867.
[3] YONOGI S, MATSUDA S, KAWAI T, et al. BEC, a novel enterotoxin of Clostridium perfringens found in human clinical isolates from acute gastroenteritis outbreaks[J]. Infection & Immunity, 2014, 82(6):2390-2399.
[4] STILES B G, KISHA P, FLEMING J M, et al. Clostridium and Bacillus binary enterotoxins: Bad for the bowels, and eukaryotic being[J]. Toxins, 2014, 6(9):2626-2656.
[5] GURJAR A A, YENNAWAR N H, YENNAWAR H P, et al. Expression, crystallization and preliminary X-ray diffraction studies of recombinant Clostridium perfringens β2-toxin[J]. Acta Crystallographica Section F, 2007, 63(6): 484-487.
[6] NAGAHAMA M, OTSUKA A, SAKURAI J. Role of tyrosine-57 and -65 in membrane-damaging and sphingomyelinase activities of Clostridium perfringens alpha-toxin[J]. Biochimica Et Biophysica Acta, 2006, 1762(1):110-114.
[7] STEVENS D L, TITBALL R W, JEPSON M, et al. Immunization with the C-domain of α-toxin prevents lethal infection, localizes tissue injury, and promotes host response to challenge with Clostridium perfringens[J]. Journal of Infectious Diseases, 2004, 190(4): 767-773.
[8] NAGAHAMA M, IIDA H, NISHIOKA E, et al. Roles of the carboxy-terminal region of Clostridium perfringens alpha toxin[J]. FEMS Microbiology Letters, 1994, 120(3):297-301.
[9] NAGAHAMA M, SAKURAI J. Threonine-74 is a key site for the activity of Clostridium perfringens alpha-toxin[J]. Microbiology and Immunology, 1996, 40(3):189-193.
[10] TITBALL R W, HUNTER S E, MARTIN K L, et al. Molecular cloning and nucleotide sequence of the alpha-toxin (phospholipase C) of Clostridium perfringens[J]. Infection & Immunity, 1989, 57(2):367-376.
[11] SAMBROOK J, FRITSCH E F, MANIATIS T. Molecular cloning[M]. 2nd Ed. New York: Cold Spring Harbor Laboratory Press, 1989: 1-50.
[12] GEOURJON C, DELÉAGE G. SOPMA: significant improvements in protein secondary structure prediction by consensus prediction from multiple alignments[J]. Bioinformatics, 1995, 77(6):681-684.
[13] ARNOLD K, BORDOLI L, KOPP J, et al. The SWISS-MODEL workspace: a web-based environment for protein structure homology modelling[J]. Bioinformatics, 2006, 22(2):195-201.
[14] 宫语晨, 许崇利, 钱爱东, 等. A型魏氏梭菌α毒素Asp-56基因定点突变与结构分析[J]. 中国兽药杂志, 2014, 48(6):1-7.
GONG Y C, XU C L, QIAN A D, et al. Site-directed mutagenesis and structure analysis of alpha-toxin gene Asp-56 from Clostridium welchii type A[J]. Chinese Journal of Veterinary Drug, 2014, 48(6): 1-7. (in Chinese with English abstract)
[15] 许崇波, 樛跃, 许崇利, 等. A型产气荚膜梭菌α毒素His-68基因的定点突变与结构分析[J]. 中国兽医科学, 2010, 40(8):811-816.
XU C B, PANG Y, XU C L, et al. Site-directed mutagenesis and structure analysis of Clostridium perfringens type A alpha-toxin gene His-68[J]. Chinese Veterinary Science, 2010, 40(8): 811-816. (in Chinese with English abstract)
[16] 樛跃, 许崇波. A型产气荚膜梭菌α毒素Thr-272基因定点突变与结构分析[J].生物技术通报,2012 (10): 186-192.
PANG Y, XU C B. Site-directed mutagenesis and structure analysis of Clostridium perfringens alpha-toxin gene Thr-272[J]. Biotechnology Bulletin, 2012 (10): 186-192. (in Chinese with English abstract)
[17] 许崇波, 许崇利, 赵志军. A型产气荚膜梭菌α毒素基因表达及其免疫保护作用的初步研究[J]. 微生物学报, 2006, 46(4): 624-628.
XU C B, XU C L, ZHAO Z J. Expression of alpha-toxin gene of Clostridium perfringens type A and its primary immunological protective function[J]. Acta Microbiologica Sinica, 2006, 46(4): 624-628. (in Chinese with English abstract)

Structural analysis and identification of biological activity of alpha-toxin amino-terminal PLC from Clostridium welchii type A

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