TY - JOUR
T1 - Purification and cytotoxic properties of Bacillus cereus hemolysin II
AU - Andreeva, Zhanna I.
AU - Nesterenko, Vladimir F.
AU - Yurkov, Igor S.
AU - Budarina, Zhanna I.
AU - Sineva, Elena V.
AU - Solonin, Alexander S.
N1 - Funding Information:
This work was supported by grants from the Russian Foundation for Basic Research (03-04-48623 and 04-04-49693) and by a subgrant with Rutgers University, Office of Research and Sponsored Programs, under the Sponsor Award No. 0853 from the Burroughs Wellcome. We thank Mr. Oleg Kovalevskiy for critical reading of the manuscript and Mr. Vladimir Mikoyan for language corrections.
Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2006/5
Y1 - 2006/5
N2 - The hemolysin II from Bacillus cereus, HlyII, is a member of the β-barrel pore-forming toxin family of secreted microbial proteins that includes the Staphylococcus aureus α-toxin. Compared with other proteins of the family, hemolysin II has 90 extra amino acids at its C-terminus. To examine more closely the cytotoxic and pore-forming properties of the protein, we have cloned and expressed it in Escherichia coli. We developed a purification procedure for the matured HlyII protein from both culture media and cell extracts using a combination of cation exchange and affinity chromatography together with gel-filtration. In both cases, the fully processed HlyII protein was purified as confirmed by N-terminal sequence analysis. The HlyII protein exhibits cytolytic activity of different extent on erythrocytes from various kinds of mammals. The results presented here show for the first time that two types of human cells are sensitive to HlyII action. In view of its broad cytotoxic activity as well as the ability to interact with artificial membranes, we assume that HlyII needs no specific receptor to bind to cell membranes.
AB - The hemolysin II from Bacillus cereus, HlyII, is a member of the β-barrel pore-forming toxin family of secreted microbial proteins that includes the Staphylococcus aureus α-toxin. Compared with other proteins of the family, hemolysin II has 90 extra amino acids at its C-terminus. To examine more closely the cytotoxic and pore-forming properties of the protein, we have cloned and expressed it in Escherichia coli. We developed a purification procedure for the matured HlyII protein from both culture media and cell extracts using a combination of cation exchange and affinity chromatography together with gel-filtration. In both cases, the fully processed HlyII protein was purified as confirmed by N-terminal sequence analysis. The HlyII protein exhibits cytolytic activity of different extent on erythrocytes from various kinds of mammals. The results presented here show for the first time that two types of human cells are sensitive to HlyII action. In view of its broad cytotoxic activity as well as the ability to interact with artificial membranes, we assume that HlyII needs no specific receptor to bind to cell membranes.
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U2 - 10.1016/j.pep.2005.10.030
DO - 10.1016/j.pep.2005.10.030
M3 - Article
C2 - 16380268
AN - SCOPUS:33646080106
SN - 1046-5928
VL - 47
SP - 186
EP - 193
JO - Protein Expression and Purification
JF - Protein Expression and Purification
IS - 1
ER -