دورية أكاديمية
Nanomolar Binding of an Antibiotic Peptide to DNA Measured with Raman Spectroscopy.
| العنوان: | Nanomolar Binding of an Antibiotic Peptide to DNA Measured with Raman Spectroscopy. |
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| المؤلفون: | Myres GJ; Department of Chemistry, University of Utah, 315 South 1400 East, Salt Lake City, Utah 84112-0850 United States., Harris JM; Department of Chemistry, University of Utah, 315 South 1400 East, Salt Lake City, Utah 84112-0850 United States. |
| المصدر: | Langmuir : the ACS journal of surfaces and colloids [Langmuir] 2023 Mar 21; Vol. 39 (11), pp. 4150-4160. Date of Electronic Publication: 2023 Mar 08. |
| نوع المنشور: | Journal Article; Research Support, U.S. Gov't, Non-P.H.S. |
| اللغة: | English |
| بيانات الدورية: | Publisher: American Chemical Society Country of Publication: United States NLM ID: 9882736 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1520-5827 (Electronic) Linking ISSN: 07437463 NLM ISO Abbreviation: Langmuir Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Washington, DC : American Chemical Society, c1985- |
| مواضيع طبية MeSH: | Spectrum Analysis, Raman* , Netropsin*/chemistry , Netropsin*/metabolism, Base Sequence ; Nucleic Acid Conformation ; DNA/chemistry ; Binding Sites ; Anti-Bacterial Agents |
| مستخلص: | Immobilization of DNA to surfaces offers a convenient means of screening the binding affinity and selectivity of potential small-molecule therapeutic candidates. Unfortunately, most surface-sensitive methods for detecting these binding interactions are not informative of the molecular structure, information that is valuable for understanding the non-covalent interactions that stabilize binding. In this work, we report a method to meet this challenge by employing confocal Raman microscopy to quantify the association of a minor-groove-binding antimicrobial peptide, netropsin, to duplex DNA hairpin sequences immobilized on the interior surfaces of porous silica particles. To assess binding selectivity, particles functionalized with different sequences of DNA were equilibrated with solutions of 100 nM netropsin, and selective association was detected based on the presence of netropsin Raman scattering in the particles. The selectivity study revealed that netropsin binds to sequences of duplex DNA having AT-rich recognition regions. To quantify binding affinities, these AT-rich DNA sequences were equilibrated with a range of netropsin solution concentrations (1 to 100 nM). Raman scattering intensities of netropsin versus solution concentration were well described by single-binding-site Langmuir isotherms with nanomolar dissociation constants, in agreement with previous isothermal calorimetry and surface plasmon resonance results. Target sequence binding was accompanied with changes in netropsin and DNA vibrational modes consistent with the hydrogen bonding between the amide groups of netropsin and adenine and thymine bases in the DNA minor groove. The binding of netropsin to a control sequence lacking the AT-rich recognition region exhibited an affinity nearly 4 orders of magnitude weaker than found for the target sequences. The Raman spectrum of netropsin interacting with this control sequence showed broad pyrrole and amide mode vibrations at frequencies similar to a free solution, revealing less constrained conformations compared with the specific binding interactions observed with AT-rich sequences. |
| المشرفين على المادة: | 64B3O0RD7N (Netropsin) 9007-49-2 (DNA) 0 (Anti-Bacterial Agents) |
| تواريخ الأحداث: | Date Created: 20230308 Date Completed: 20230322 Latest Revision: 20230412 |
| رمز التحديث: | 20231215 |
| DOI: | 10.1021/acs.langmuir.3c00099 |
| PMID: | 36888905 |
| قاعدة البيانات: | MEDLINE |
Find this issue from the American Chemical Society | تدمد: | 1520-5827 |
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| DOI: | 10.1021/acs.langmuir.3c00099 |