BioSAXS Measurements Reveal That Two Antimicrobial Peptides Induce Similar Molecular Changes in Gram-Negative and Gram-Positive Bacteria
Two highly active short broad-spectrum AMPs (14D and 69D) with unknown mode of action have been investigated in regards to their effect against the Gram-negative bacteria Escherichia coli and the Gram-positive bacteria methicillin-resistant Staphylococcus aureus (MRSA). Minimal inhibitory concentrat...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2019-09-01
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| Series: | Frontiers in Pharmacology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fphar.2019.01127/full |
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| author | Andreas von Gundlach Martin P. Ashby Jurnorain Gani Paula Matilde Lopez-Perez Alan Roy Cookson Sharon Ann Huws Christoph Rumancev Vasil M. Garamus Ralf Mikut Axel Rosenhahn Kai Hilpert |
| author_facet | Andreas von Gundlach Martin P. Ashby Jurnorain Gani Paula Matilde Lopez-Perez Alan Roy Cookson Sharon Ann Huws Christoph Rumancev Vasil M. Garamus Ralf Mikut Axel Rosenhahn Kai Hilpert |
| author_sort | Andreas von Gundlach |
| collection | DOAJ |
| description | Two highly active short broad-spectrum AMPs (14D and 69D) with unknown mode of action have been investigated in regards to their effect against the Gram-negative bacteria Escherichia coli and the Gram-positive bacteria methicillin-resistant Staphylococcus aureus (MRSA). Minimal inhibitory concentration (MIC) measurements using a cell density of 108 cfu/ml resulted in values between 16 and 32 µg/ml. Time-kill experiments using 108 cfu/ml revealed complete killing, except for 69D in combination with MRSA, where bacterial load was reduced a million times. Small-angle X-ray scattering of biological samples (BioSAXS) at 108 cfu/ml was applied to investigate the ultrastructural changes in E. coli and MRSA in response to these two broad-spectrum AMPs. In addition, electron microscopy (EM) was performed to visualize the treated and non-treated bacteria. As expected, the scattering curves generated using BioSAXS show the ultrastructure of the Gram-positive and Gram-negative bacteria to be very different (BioSAXS is not susceptible to the outer shape). After treatment with either peptide, the scattering curves of E. coli and MRSA cells are much more alike. Whereas in EM, it is notoriously difficult to observe changes for spherical Gram-positives; the BioSAXS results are superior and reveal strongly similar effects for both peptides induced in Gram-positive as well as Gram-negative bacteria. Given the high-throughput possibility and robust statistics, BioSAXS can support and speed up mode of action research in AMPs and other antimicrobial compounds, making a contribution toward the development of urgently needed drugs against resistant bacteria. |
| first_indexed | 2024-12-21T07:42:22Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 1663-9812 |
| language | English |
| last_indexed | 2024-12-21T07:42:22Z |
| publishDate | 2019-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Pharmacology |
| spelling | doaj.art-fd889a1536a1414588891f2c989b26a52022-12-21T19:11:17ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122019-09-011010.3389/fphar.2019.01127466414BioSAXS Measurements Reveal That Two Antimicrobial Peptides Induce Similar Molecular Changes in Gram-Negative and Gram-Positive BacteriaAndreas von Gundlach0Martin P. Ashby1Jurnorain Gani2Paula Matilde Lopez-Perez3Alan Roy Cookson4Sharon Ann Huws5Christoph Rumancev6Vasil M. Garamus7Ralf Mikut8Axel Rosenhahn9Kai Hilpert10Analytical Chemistry - Biointerfaces, Ruhr-University Bochum, Bochum, GermanyInstitute of Infection and Immunology, St. George’s University of London (SGUL), London, United KingdomInstitute of Infection and Immunology, St. George’s University of London (SGUL), London, United KingdomTiKa Diagnostics Ltd, London, United KingdomInstitute of Biological, Environmental & Rural Sciences (IBERS), Aberystwyth University, Aberystwyth, United KingdomInstitute of Global Food Security, School of Biological Sciences, Queens University Belfast, Medical Biology Centre, Belfast, United KingdomAnalytical Chemistry - Biointerfaces, Ruhr-University Bochum, Bochum, GermanyHelmholtz-Zentrum Geesthacht, Zentrum für Material- und Küstenforschung GmbH, Geesthacht, GermanyInstitute for Automation and Applied Informatics (IAI), Karlsruhe Institute of Technology (KIT), Karlsruhe, GermanyAnalytical Chemistry - Biointerfaces, Ruhr-University Bochum, Bochum, GermanyInstitute of Infection and Immunology, St. George’s University of London (SGUL), London, United KingdomTwo highly active short broad-spectrum AMPs (14D and 69D) with unknown mode of action have been investigated in regards to their effect against the Gram-negative bacteria Escherichia coli and the Gram-positive bacteria methicillin-resistant Staphylococcus aureus (MRSA). Minimal inhibitory concentration (MIC) measurements using a cell density of 108 cfu/ml resulted in values between 16 and 32 µg/ml. Time-kill experiments using 108 cfu/ml revealed complete killing, except for 69D in combination with MRSA, where bacterial load was reduced a million times. Small-angle X-ray scattering of biological samples (BioSAXS) at 108 cfu/ml was applied to investigate the ultrastructural changes in E. coli and MRSA in response to these two broad-spectrum AMPs. In addition, electron microscopy (EM) was performed to visualize the treated and non-treated bacteria. As expected, the scattering curves generated using BioSAXS show the ultrastructure of the Gram-positive and Gram-negative bacteria to be very different (BioSAXS is not susceptible to the outer shape). After treatment with either peptide, the scattering curves of E. coli and MRSA cells are much more alike. Whereas in EM, it is notoriously difficult to observe changes for spherical Gram-positives; the BioSAXS results are superior and reveal strongly similar effects for both peptides induced in Gram-positive as well as Gram-negative bacteria. Given the high-throughput possibility and robust statistics, BioSAXS can support and speed up mode of action research in AMPs and other antimicrobial compounds, making a contribution toward the development of urgently needed drugs against resistant bacteria.https://www.frontiersin.org/article/10.3389/fphar.2019.01127/fullantimicrobial peptidemode of actionSAXSMRSAelectron microscopyE. coli |
| spellingShingle | Andreas von Gundlach Martin P. Ashby Jurnorain Gani Paula Matilde Lopez-Perez Alan Roy Cookson Sharon Ann Huws Christoph Rumancev Vasil M. Garamus Ralf Mikut Axel Rosenhahn Kai Hilpert BioSAXS Measurements Reveal That Two Antimicrobial Peptides Induce Similar Molecular Changes in Gram-Negative and Gram-Positive Bacteria Frontiers in Pharmacology antimicrobial peptide mode of action SAXS MRSA electron microscopy E. coli |
| title | BioSAXS Measurements Reveal That Two Antimicrobial Peptides Induce Similar Molecular Changes in Gram-Negative and Gram-Positive Bacteria |
| title_full | BioSAXS Measurements Reveal That Two Antimicrobial Peptides Induce Similar Molecular Changes in Gram-Negative and Gram-Positive Bacteria |
| title_fullStr | BioSAXS Measurements Reveal That Two Antimicrobial Peptides Induce Similar Molecular Changes in Gram-Negative and Gram-Positive Bacteria |
| title_full_unstemmed | BioSAXS Measurements Reveal That Two Antimicrobial Peptides Induce Similar Molecular Changes in Gram-Negative and Gram-Positive Bacteria |
| title_short | BioSAXS Measurements Reveal That Two Antimicrobial Peptides Induce Similar Molecular Changes in Gram-Negative and Gram-Positive Bacteria |
| title_sort | biosaxs measurements reveal that two antimicrobial peptides induce similar molecular changes in gram negative and gram positive bacteria |
| topic | antimicrobial peptide mode of action SAXS MRSA electron microscopy E. coli |
| url | https://www.frontiersin.org/article/10.3389/fphar.2019.01127/full |
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