Mechanism of interaction of optimized Limulus-derived cyclic peptides with endotoxins: thermodynamic, biophysical and microbiological analysis
On the basis of formerly investigated peptides corresponding to the endotoxin-binding domain from LALF [Limulus anti-LPS (lipopolysaccharide) factor], a protein from Limulus polyphemus, we have designed and synthesized peptides of different lengths with the aim of obtaining potential therapeutic age...
| Main Authors: | , , , , , , , , , |
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| Format: | info:eu-repo/semantics/article |
| Language: | eng |
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Portland Press
2013
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| Subjects: | |
| Online Access: | https://hdl.handle.net/10171/29427 |
| _version_ | 1793400008770322432 |
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| author | Andrä, J. (Jörg) Howe, J. (Jörg) Garidel, P. (Patrick) Rossle, M. (Manfred) Richter, W. (Walter) Leiva, J. (José) Moriyon, I. (Ignacio) Bartels, R. (Rainer) Gutsmann, T. (Thomas) Brandenburg, K. (Klaus) |
| author_facet | Andrä, J. (Jörg) Howe, J. (Jörg) Garidel, P. (Patrick) Rossle, M. (Manfred) Richter, W. (Walter) Leiva, J. (José) Moriyon, I. (Ignacio) Bartels, R. (Rainer) Gutsmann, T. (Thomas) Brandenburg, K. (Klaus) |
| author_sort | Andrä, J. (Jörg) |
| collection | DSpace |
| description | On the basis of formerly investigated peptides corresponding to the endotoxin-binding domain from LALF [Limulus anti-LPS (lipopolysaccharide) factor], a protein from Limulus polyphemus, we have designed and synthesized peptides of different lengths with the aim of obtaining potential therapeutic agents against septic shock syndrome. For an understanding of the mechanisms of action, we performed a detailed physicochemical and biophysical analysis of the interaction of rough mutant LPS with these peptides by applying FTIR (Fourier-transform infrared) spectroscopy, SAXS (small-angle X-ray scattering), calorimetric techniques [DSC (differential scanning calorimetry) and ITC (isothermal titration calorimetry)] and FFTEM (freeze-fracture transmission electron microscopy). Also, the action of the peptides on bacteria of different origin in microbial assays was investigated. Using FTIR and DSC, our results indicated a strong fluidization of the lipid A acyl chains due to peptide binding, with a decrease in the endothermic melting enthalpy change of the acyl chains down to a complete disappearance in the 1:0.5 to 1:2 [LPS]:[peptide] molar ratio range. Via ITC, it was deduced that the binding is a clearly exothermic process which becomes saturated at a 1:0.5 to 1:2 [LPS]:[peptide] molar ratio range. The results obtained with SAXS indicated a drastic change of the aggregate structures of LPS into a multilamellar stack, which was visualized in electron micrographs as hundreds of lamellar layers. This can be directly correlated with the inhibition of the LPS-induced production of tumour necrosis factor alpha in human mononuclear cells, but not with the action of the peptides on bacteria. |
| format | info:eu-repo/semantics/article |
| id | oai:dadun.unav.edu:10171-29427 |
| institution | Universidad de Navarra |
| language | eng |
| publishDate | 2013 |
| publisher | Portland Press |
| record_format | dspace |
| spelling | oai:dadun.unav.edu:10171-294272020-03-03T23:27:01Z Mechanism of interaction of optimized Limulus-derived cyclic peptides with endotoxins: thermodynamic, biophysical and microbiological analysis Andrä, J. (Jörg) Howe, J. (Jörg) Garidel, P. (Patrick) Rossle, M. (Manfred) Richter, W. (Walter) Leiva, J. (José) Moriyon, I. (Ignacio) Bartels, R. (Rainer) Gutsmann, T. (Thomas) Brandenburg, K. (Klaus) Biophysics Endotoxins pharmacology Invertebrate hormones chemistry Peptides Cyclic chemistry Thermodynamics On the basis of formerly investigated peptides corresponding to the endotoxin-binding domain from LALF [Limulus anti-LPS (lipopolysaccharide) factor], a protein from Limulus polyphemus, we have designed and synthesized peptides of different lengths with the aim of obtaining potential therapeutic agents against septic shock syndrome. For an understanding of the mechanisms of action, we performed a detailed physicochemical and biophysical analysis of the interaction of rough mutant LPS with these peptides by applying FTIR (Fourier-transform infrared) spectroscopy, SAXS (small-angle X-ray scattering), calorimetric techniques [DSC (differential scanning calorimetry) and ITC (isothermal titration calorimetry)] and FFTEM (freeze-fracture transmission electron microscopy). Also, the action of the peptides on bacteria of different origin in microbial assays was investigated. Using FTIR and DSC, our results indicated a strong fluidization of the lipid A acyl chains due to peptide binding, with a decrease in the endothermic melting enthalpy change of the acyl chains down to a complete disappearance in the 1:0.5 to 1:2 [LPS]:[peptide] molar ratio range. Via ITC, it was deduced that the binding is a clearly exothermic process which becomes saturated at a 1:0.5 to 1:2 [LPS]:[peptide] molar ratio range. The results obtained with SAXS indicated a drastic change of the aggregate structures of LPS into a multilamellar stack, which was visualized in electron micrographs as hundreds of lamellar layers. This can be directly correlated with the inhibition of the LPS-induced production of tumour necrosis factor alpha in human mononuclear cells, but not with the action of the peptides on bacteria. 2013-06-25T08:15:56Z 2013-06-25T08:15:56Z 2007 info:eu-repo/semantics/article https://hdl.handle.net/10171/29427 eng info:eu-repo/semantics/openAccess application/pdf Portland Press |
| spellingShingle | Biophysics Endotoxins pharmacology Invertebrate hormones chemistry Peptides Cyclic chemistry Thermodynamics Andrä, J. (Jörg) Howe, J. (Jörg) Garidel, P. (Patrick) Rossle, M. (Manfred) Richter, W. (Walter) Leiva, J. (José) Moriyon, I. (Ignacio) Bartels, R. (Rainer) Gutsmann, T. (Thomas) Brandenburg, K. (Klaus) Mechanism of interaction of optimized Limulus-derived cyclic peptides with endotoxins: thermodynamic, biophysical and microbiological analysis |
| title | Mechanism of interaction of optimized Limulus-derived cyclic peptides with endotoxins: thermodynamic, biophysical and microbiological analysis |
| title_full | Mechanism of interaction of optimized Limulus-derived cyclic peptides with endotoxins: thermodynamic, biophysical and microbiological analysis |
| title_fullStr | Mechanism of interaction of optimized Limulus-derived cyclic peptides with endotoxins: thermodynamic, biophysical and microbiological analysis |
| title_full_unstemmed | Mechanism of interaction of optimized Limulus-derived cyclic peptides with endotoxins: thermodynamic, biophysical and microbiological analysis |
| title_short | Mechanism of interaction of optimized Limulus-derived cyclic peptides with endotoxins: thermodynamic, biophysical and microbiological analysis |
| title_sort | mechanism of interaction of optimized limulus-derived cyclic peptides with endotoxins: thermodynamic, biophysical and microbiological analysis |
| topic | Biophysics Endotoxins pharmacology Invertebrate hormones chemistry Peptides Cyclic chemistry Thermodynamics |
| url | https://hdl.handle.net/10171/29427 |
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