Linear Dichroism Measurements for the Study of Protein-DNA Interactions
Linear dichroism (LD) is a differential polarized light absorption spectroscopy used for studying filamentous molecules such as DNA and protein filaments. In this study, we review the applications of LD for the analysis of DNA-protein interactions. LD signals can be measured in a solution by alignin...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-11-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/22/16092 |
| _version_ | 1797459046940278784 |
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| author | Masayuki Takahashi Bengt Norden |
| author_facet | Masayuki Takahashi Bengt Norden |
| author_sort | Masayuki Takahashi |
| collection | DOAJ |
| description | Linear dichroism (LD) is a differential polarized light absorption spectroscopy used for studying filamentous molecules such as DNA and protein filaments. In this study, we review the applications of LD for the analysis of DNA-protein interactions. LD signals can be measured in a solution by aligning the sample using flow-induced shear force or a strong electric field. The signal generated is related to the local orientation of chromophores, such as DNA bases, relative to the filament axis. LD can thus assess the tilt and roll of DNA bases and distinguish intercalating from groove-binding ligands. The intensity of the LD signal depends upon the degree of macroscopic orientation. Therefore, DNA shortening and bending can be detected by a decrease in LD signal intensity. As examples of LD applications, we present a kinetic study of DNA digestion by restriction enzymes and structural analyses of homologous recombination intermediates, i.e., RecA and Rad51 recombinase complexes with single-stranded DNA. LD shows that the DNA bases in these complexes are preferentially oriented perpendicular to the filament axis only in the presence of activators, suggesting the importance of organized base orientation for the reaction. LD measurements detect DNA bending by the CRP transcription activator protein, as well as by the UvrB DNA repair protein. LD can thus provide information about the structures of protein-DNA complexes under various conditions and in real time. |
| first_indexed | 2024-03-09T16:46:48Z |
| format | Article |
| id | doaj.art-43a473d2ab3e4c47a66bdce54241cdf6 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T16:46:48Z |
| publishDate | 2023-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-43a473d2ab3e4c47a66bdce54241cdf62023-11-24T14:45:52ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-11-0124221609210.3390/ijms242216092Linear Dichroism Measurements for the Study of Protein-DNA InteractionsMasayuki Takahashi0Bengt Norden1School of Life Science and Technology, Tokyo Institute of Technology, Oookayama, Meguro, Tokyo 152-8550, JapanDepartment of Chemical and Biological Engineering, Chemistry, Chalmers University of Technology, 412 96 Gothenburg, SwedenLinear dichroism (LD) is a differential polarized light absorption spectroscopy used for studying filamentous molecules such as DNA and protein filaments. In this study, we review the applications of LD for the analysis of DNA-protein interactions. LD signals can be measured in a solution by aligning the sample using flow-induced shear force or a strong electric field. The signal generated is related to the local orientation of chromophores, such as DNA bases, relative to the filament axis. LD can thus assess the tilt and roll of DNA bases and distinguish intercalating from groove-binding ligands. The intensity of the LD signal depends upon the degree of macroscopic orientation. Therefore, DNA shortening and bending can be detected by a decrease in LD signal intensity. As examples of LD applications, we present a kinetic study of DNA digestion by restriction enzymes and structural analyses of homologous recombination intermediates, i.e., RecA and Rad51 recombinase complexes with single-stranded DNA. LD shows that the DNA bases in these complexes are preferentially oriented perpendicular to the filament axis only in the presence of activators, suggesting the importance of organized base orientation for the reaction. LD measurements detect DNA bending by the CRP transcription activator protein, as well as by the UvrB DNA repair protein. LD can thus provide information about the structures of protein-DNA complexes under various conditions and in real time.https://www.mdpi.com/1422-0067/24/22/16092linear dichroism (LD)DNA/protein complexRecARad51homologous recombinationcyclic AMP receptor protein (CRP) |
| spellingShingle | Masayuki Takahashi Bengt Norden Linear Dichroism Measurements for the Study of Protein-DNA Interactions International Journal of Molecular Sciences linear dichroism (LD) DNA/protein complex RecA Rad51 homologous recombination cyclic AMP receptor protein (CRP) |
| title | Linear Dichroism Measurements for the Study of Protein-DNA Interactions |
| title_full | Linear Dichroism Measurements for the Study of Protein-DNA Interactions |
| title_fullStr | Linear Dichroism Measurements for the Study of Protein-DNA Interactions |
| title_full_unstemmed | Linear Dichroism Measurements for the Study of Protein-DNA Interactions |
| title_short | Linear Dichroism Measurements for the Study of Protein-DNA Interactions |
| title_sort | linear dichroism measurements for the study of protein dna interactions |
| topic | linear dichroism (LD) DNA/protein complex RecA Rad51 homologous recombination cyclic AMP receptor protein (CRP) |
| url | https://www.mdpi.com/1422-0067/24/22/16092 |
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