Protein-Lipid Interaction of Cytolytic Toxin Cyt2Aa2 on Model Lipid Bilayers of Erythrocyte Cell Membrane

被引:3
|
作者
Tharad, Sudarat [1 ]
Promdonkoy, Boonhiang [2 ]
Toca-Herrera, Jose L. [1 ]
机构
[1] Univ Nat Resources & Life Sci BOKU, Dept Nanobiotechnol, Inst Biophys, A-1190 Vienna, Austria
[2] Natl Sci & Technol Dev Agcy, Natl Ctr Genet Engn & Biotechnol, Pathum Thani 12120, Thailand
基金
奥地利科学基金会;
关键词
Cyt2Aa2; toxin; protein-lipid binding; erythrocyte membrane; AFM; QCM-D; THURINGIENSIS SUBSP ISRAELENSIS; DELTA-ENDOTOXIN; MECHANISM; CRYSTAL; TOXICITY; REVEALS; BINDING; INSECT; CYT1A; LOOP;
D O I
10.3390/toxins12040226
中图分类号
TS2 [食品工业];
学科分类号
0832 ;
摘要
Cytolytic toxin (Cyt) is a toxin among Bacillus thuringiensis insecticidal proteins. Cyt toxin directly interacts with membrane lipids for cytolytic action. However, low hemolytic activity is desired to avoid non-specific effects in mammals. In this work, the interaction between Cyt2Aa2 toxin and model lipid bilayers mimicking the erythrocyte membrane was investigated for Cyt2Aa2 wild type (WT) and the T144A mutant, a variant with lower hemolytic activity. Quartz crystal microbalance with dissipation (QCM-D) results revealed a smaller lipid binding capacity for the T144A mutant than for the WT. In particular, the T144A mutant was unable to bind to the phosphatidylcholine lipid (POPC) bilayer. However, the addition of cholesterol (Chol) or sphingomyelin (SM) to the POPC bilayer promoted binding of the T144 mutant. Moreover, atomic force microscopy (AFM) images unveiled small aggregates of the T144A mutant on the 1:1 sphingomyelin/POPC bilayers. In contrast, the lipid binding trend for WT and T144A mutant was comparable for the 1:0.4 POPC/cholesterol and the 1:1:1 sphingomyelin/POPC/cholesterol bilayers. Furthermore, the binding of WT and T144A mutant onto erythrocyte cells was investigated. The experiments showed that the T144A mutant and the WT bind onto different areas of the erythrocyte membrane. Overall the results suggest that the T144 residue plays an important role for lipid binding.
引用
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页数:12
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