2v3l
From Proteopedia
(New page: 200px<br /><applet load="2v3l" size="350" color="white" frame="true" align="right" spinBox="true" caption="2v3l" /> '''ORIENTATIONAL AND DYNAMICAL HETEROGENEITY OF...) |
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==Overview== | ==Overview== | ||
| - | The comparison of Forster resonance energy transfer (FRET) efficiencies | + | The comparison of Forster resonance energy transfer (FRET) efficiencies between two fluorophores covalently attached to a single protein or DNA molecule is an elegant approach for deducing information about their structural and dynamical heterogeneity. For a more detailed structural interpretation of single-molecule FRET assays, information about the positions as well as the dynamics of the dye labels attached to the biomolecule is important. In this work, Rhodamine 6G (2-[3'-(ethylamino)-6'-(ethylimino)-2',7'-dimethyl-6'H-xanthen-9'-yl]-benz oic acid) bound to the 5'-end of a 20 base pair long DNA duplex is investigated by both single-molecule multiparameter fluorescence detection (MFD) experiments and NMR spectroscopy. Rhodamine 6G is commonly employed in nucleic acid research as a FRET dye. MFD experiments directly reveal the equilibrium of the dye bound to DNA between three heterogeneous environments, which are characterized by distinct fluorescence lifetime and intensity distributions as a result of different guanine-dye excited-state electron transfer interactions. Sub-ensemble fluorescence autocorrelation analysis shows the highly dynamic character of the dye-DNA interactions ranging from nano- to milliseconds and species-specific triplet relaxation times. Two-dimensional NMR spectroscopy corroborates this information by the determination of the detailed geometric structures of the dye-nucleobase complex and their assignment to each population observed in the single-molecule fluorescence experiments. From both methods, a consistent and detailed molecular description of the structural and dynamical heterogeneity is obtained. |
==About this Structure== | ==About this Structure== | ||
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==Reference== | ==Reference== | ||
| - | Orientational and | + | Orientational and dynamical heterogeneity of rhodamine 6G terminally attached to a DNA helix revealed by NMR and single-molecule fluorescence spectroscopy., Neubauer H, Gaiko N, Berger S, Schaffer J, Eggeling C, Tuma J, Verdier L, Seidel CA, Griesinger C, Volkmer A, J Am Chem Soc. 2007 Oct 24;129(42):12746-55. Epub 2007 Sep 27. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=17900110 17900110] |
[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Berger, S.]] | [[Category: Berger, S.]] | ||
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[[Category: Neubauer, H.]] | [[Category: Neubauer, H.]] | ||
[[Category: Schaffer, J.]] | [[Category: Schaffer, J.]] | ||
| - | [[Category: Seidel, C | + | [[Category: Seidel, C A.M.]] |
[[Category: Tuma, J.]] | [[Category: Tuma, J.]] | ||
[[Category: Verdier, L.]] | [[Category: Verdier, L.]] | ||
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[[Category: nucleic acid]] | [[Category: nucleic acid]] | ||
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 18:52:57 2008'' |
Revision as of 16:52, 21 February 2008
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ORIENTATIONAL AND DYNAMICAL HETEROGENEITY OF RHODAMINE 6G TERMINALLY ATTACHED TO A DNA HELIX
Overview
The comparison of Forster resonance energy transfer (FRET) efficiencies between two fluorophores covalently attached to a single protein or DNA molecule is an elegant approach for deducing information about their structural and dynamical heterogeneity. For a more detailed structural interpretation of single-molecule FRET assays, information about the positions as well as the dynamics of the dye labels attached to the biomolecule is important. In this work, Rhodamine 6G (2-[3'-(ethylamino)-6'-(ethylimino)-2',7'-dimethyl-6'H-xanthen-9'-yl]-benz oic acid) bound to the 5'-end of a 20 base pair long DNA duplex is investigated by both single-molecule multiparameter fluorescence detection (MFD) experiments and NMR spectroscopy. Rhodamine 6G is commonly employed in nucleic acid research as a FRET dye. MFD experiments directly reveal the equilibrium of the dye bound to DNA between three heterogeneous environments, which are characterized by distinct fluorescence lifetime and intensity distributions as a result of different guanine-dye excited-state electron transfer interactions. Sub-ensemble fluorescence autocorrelation analysis shows the highly dynamic character of the dye-DNA interactions ranging from nano- to milliseconds and species-specific triplet relaxation times. Two-dimensional NMR spectroscopy corroborates this information by the determination of the detailed geometric structures of the dye-nucleobase complex and their assignment to each population observed in the single-molecule fluorescence experiments. From both methods, a consistent and detailed molecular description of the structural and dynamical heterogeneity is obtained.
About this Structure
2V3L is a Single protein structure of sequence from [1] with as ligand. Full crystallographic information is available from OCA.
Reference
Orientational and dynamical heterogeneity of rhodamine 6G terminally attached to a DNA helix revealed by NMR and single-molecule fluorescence spectroscopy., Neubauer H, Gaiko N, Berger S, Schaffer J, Eggeling C, Tuma J, Verdier L, Seidel CA, Griesinger C, Volkmer A, J Am Chem Soc. 2007 Oct 24;129(42):12746-55. Epub 2007 Sep 27. PMID:17900110
Page seeded by OCA on Thu Feb 21 18:52:57 2008
Categories: Single protein | Berger, S. | Eggeling, C. | Gaiko, N. | Griesinger, C. | Neubauer, H. | Schaffer, J. | Seidel, C A.M. | Tuma, J. | Verdier, L. | Volkmer, A. | R6G | Dna | Nucleic acid
