A Monoclonal Antibody against the C-Terminal Domain of <i>Bacillus cereus</i> Hemolysin II Inhibits HlyII Cytolytic Activity
<i>Bacillus cereus</i> is the fourth most common cause of foodborne illnesses that produces a variety of pore-forming proteins as the main pathogenic factors. <i>B. cereus</i> hemolysin II (HlyII), belonging to pore-forming β-barrel toxins, has a C-terminal extension of 94 am...
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| Format: | Article |
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MDPI AG
2020-12-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/12/12/806 |
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| author | Natalia Rudenko Alexey Nagel Anna Zamyatina Anna Karatovskaya Vadim Salyamov Zhanna Andreeva-Kovalevskaya Alexander Siunov Alexander Kolesnikov Anna Shepelyakovskaya Khanafiy Boziev Bogdan Melnik Fedor Brovko Alexander Solonin |
| author_facet | Natalia Rudenko Alexey Nagel Anna Zamyatina Anna Karatovskaya Vadim Salyamov Zhanna Andreeva-Kovalevskaya Alexander Siunov Alexander Kolesnikov Anna Shepelyakovskaya Khanafiy Boziev Bogdan Melnik Fedor Brovko Alexander Solonin |
| author_sort | Natalia Rudenko |
| collection | DOAJ |
| description | <i>Bacillus cereus</i> is the fourth most common cause of foodborne illnesses that produces a variety of pore-forming proteins as the main pathogenic factors. <i>B. cereus</i> hemolysin II (HlyII), belonging to pore-forming β-barrel toxins, has a C-terminal extension of 94 amino acid residues designated as HlyIICTD. An analysis of a panel of monoclonal antibodies to the recombinant HlyIICTD protein revealed the ability of the antibody HlyIIC-20 to inhibit HlyII hemolysis. A conformational epitope recognized by HlyIIC-20 was found. by the method of peptide phage display and found that it is localized in the N-terminal part of HlyIICTD. The HlyIIC-20 interacted with a monomeric form of HlyII, thus suppressing maturation of the HlyII toxin. Protection efficiencies of various <i>B. cereus</i> strains against HlyII were different and depended on the epitope amino acid composition, as well as, insignificantly, on downstream amino acids. Substitution of L324P and P324L in the hemolysins ATCC14579<sup>T</sup> and B771, respectively, determined the role of leucine localized to the epitope in suppressing the hemolysis by the antibody. Pre-incubation of HlyIIC-20 with HlyII prevented the death of mice up to an equimolar ratio. A strategy of detecting and neutralizing the toxic activity of HlyII could provide a tool for monitoring and reducing <i>B. cereus</i> pathogenicity. |
| first_indexed | 2024-03-10T13:55:10Z |
| format | Article |
| id | doaj.art-6120877331f84866842af14a4d9bb73f |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-10T13:55:10Z |
| publishDate | 2020-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-6120877331f84866842af14a4d9bb73f2023-11-21T01:41:11ZengMDPI AGToxins2072-66512020-12-01121280610.3390/toxins12120806A Monoclonal Antibody against the C-Terminal Domain of <i>Bacillus cereus</i> Hemolysin II Inhibits HlyII Cytolytic ActivityNatalia Rudenko0Alexey Nagel1Anna Zamyatina2Anna Karatovskaya3Vadim Salyamov4Zhanna Andreeva-Kovalevskaya5Alexander Siunov6Alexander Kolesnikov7Anna Shepelyakovskaya8Khanafiy Boziev9Bogdan Melnik10Fedor Brovko11Alexander Solonin12Pushchino Branch, Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 6 Prospekt Nauki, 142290 Pushchino, Moscow Region, RussiaFSBIS FRC Pushchino Scientific Centre of Biological Research, G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, 5 Prospekt Nauki, 142290 Pushchino, Moscow Region, RussiaPushchino Branch, Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 6 Prospekt Nauki, 142290 Pushchino, Moscow Region, RussiaPushchino Branch, Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 6 Prospekt Nauki, 142290 Pushchino, Moscow Region, RussiaFSBIS FRC Pushchino Scientific Centre of Biological Research, G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, 5 Prospekt Nauki, 142290 Pushchino, Moscow Region, RussiaFSBIS FRC Pushchino Scientific Centre of Biological Research, G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, 5 Prospekt Nauki, 142290 Pushchino, Moscow Region, RussiaFSBIS FRC Pushchino Scientific Centre of Biological Research, G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, 5 Prospekt Nauki, 142290 Pushchino, Moscow Region, RussiaFSBIS FRC Pushchino Scientific Centre of Biological Research, G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, 5 Prospekt Nauki, 142290 Pushchino, Moscow Region, RussiaPushchino Branch, Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 6 Prospekt Nauki, 142290 Pushchino, Moscow Region, RussiaPushchino Branch, Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 6 Prospekt Nauki, 142290 Pushchino, Moscow Region, RussiaProtein Institute of the Russian Academy of Sciences, 4 Prospekt Nauki, 142290 Pushchino, Moscow Region, RussiaPushchino Branch, Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 6 Prospekt Nauki, 142290 Pushchino, Moscow Region, RussiaFSBIS FRC Pushchino Scientific Centre of Biological Research, G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, 5 Prospekt Nauki, 142290 Pushchino, Moscow Region, Russia<i>Bacillus cereus</i> is the fourth most common cause of foodborne illnesses that produces a variety of pore-forming proteins as the main pathogenic factors. <i>B. cereus</i> hemolysin II (HlyII), belonging to pore-forming β-barrel toxins, has a C-terminal extension of 94 amino acid residues designated as HlyIICTD. An analysis of a panel of monoclonal antibodies to the recombinant HlyIICTD protein revealed the ability of the antibody HlyIIC-20 to inhibit HlyII hemolysis. A conformational epitope recognized by HlyIIC-20 was found. by the method of peptide phage display and found that it is localized in the N-terminal part of HlyIICTD. The HlyIIC-20 interacted with a monomeric form of HlyII, thus suppressing maturation of the HlyII toxin. Protection efficiencies of various <i>B. cereus</i> strains against HlyII were different and depended on the epitope amino acid composition, as well as, insignificantly, on downstream amino acids. Substitution of L324P and P324L in the hemolysins ATCC14579<sup>T</sup> and B771, respectively, determined the role of leucine localized to the epitope in suppressing the hemolysis by the antibody. Pre-incubation of HlyIIC-20 with HlyII prevented the death of mice up to an equimolar ratio. A strategy of detecting and neutralizing the toxic activity of HlyII could provide a tool for monitoring and reducing <i>B. cereus</i> pathogenicity.https://www.mdpi.com/2072-6651/12/12/806pore-forming toxinhybridomabacteriophage displayepitope mappingneutralizing monoclonal antibodyoligomerization |
| spellingShingle | Natalia Rudenko Alexey Nagel Anna Zamyatina Anna Karatovskaya Vadim Salyamov Zhanna Andreeva-Kovalevskaya Alexander Siunov Alexander Kolesnikov Anna Shepelyakovskaya Khanafiy Boziev Bogdan Melnik Fedor Brovko Alexander Solonin A Monoclonal Antibody against the C-Terminal Domain of <i>Bacillus cereus</i> Hemolysin II Inhibits HlyII Cytolytic Activity Toxins pore-forming toxin hybridoma bacteriophage display epitope mapping neutralizing monoclonal antibody oligomerization |
| title | A Monoclonal Antibody against the C-Terminal Domain of <i>Bacillus cereus</i> Hemolysin II Inhibits HlyII Cytolytic Activity |
| title_full | A Monoclonal Antibody against the C-Terminal Domain of <i>Bacillus cereus</i> Hemolysin II Inhibits HlyII Cytolytic Activity |
| title_fullStr | A Monoclonal Antibody against the C-Terminal Domain of <i>Bacillus cereus</i> Hemolysin II Inhibits HlyII Cytolytic Activity |
| title_full_unstemmed | A Monoclonal Antibody against the C-Terminal Domain of <i>Bacillus cereus</i> Hemolysin II Inhibits HlyII Cytolytic Activity |
| title_short | A Monoclonal Antibody against the C-Terminal Domain of <i>Bacillus cereus</i> Hemolysin II Inhibits HlyII Cytolytic Activity |
| title_sort | monoclonal antibody against the c terminal domain of i bacillus cereus i hemolysin ii inhibits hlyii cytolytic activity |
| topic | pore-forming toxin hybridoma bacteriophage display epitope mapping neutralizing monoclonal antibody oligomerization |
| url | https://www.mdpi.com/2072-6651/12/12/806 |
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