1aa3
From Proteopedia
(New page: 200px<br /><applet load="1aa3" size="450" color="white" frame="true" align="right" spinBox="true" caption="1aa3" /> '''C-TERMINAL DOMAIN OF THE E. COLI RECA, NMR, ...) |
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| - | [[Image:1aa3.gif|left|200px]]<br /><applet load="1aa3" size=" | + | [[Image:1aa3.gif|left|200px]]<br /><applet load="1aa3" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1aa3" /> | caption="1aa3" /> | ||
'''C-TERMINAL DOMAIN OF THE E. COLI RECA, NMR, MINIMIZED AVERAGE STRUCTURE'''<br /> | '''C-TERMINAL DOMAIN OF THE E. COLI RECA, NMR, MINIMIZED AVERAGE STRUCTURE'''<br /> | ||
==Overview== | ==Overview== | ||
| - | RecA protein and its homologs catalyze homologous pairing of dsDNA and | + | RecA protein and its homologs catalyze homologous pairing of dsDNA and ssDNA, a critical reaction in homologous genetic recombination in various organisms from a virus, microbes to higher eukaryotes. In this reaction, RecA protein forms a nucleoprotein filament on ssDNA, which in turn binds to naked dsDNA for homology search. We suggested that the C-terminal domain of RecA protein plays a role in capturing the dsDNA. Here, we isolated the C-terminal domain as a soluble form and determined the solution structure by NMR spectroscopy. The overall folding of the NMR structure agrees with that of the corresponding part of the reported crystal structure, but a remarkable difference was found in a solvent-exposed region due to intermolecular contacts in the crystal. Then, we studied the interaction between the C-terminal domain and DNA, and found that significant chemical shift changes were induced in a specific region by titration with dsDNA. SsDNA induced a much smaller chemical shift perturbation. The difference of DNA concentrations to give the half-saturation of the chemical shift change showed a higher affinity of the C-terminal region toward dsDNA. Combined with our previous results, these provide direct evidence that the defined region in the C-terminal domain furnishes a binding surface for DNA. |
==About this Structure== | ==About this Structure== | ||
| - | 1AA3 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli]. Full crystallographic information is available from [http:// | + | 1AA3 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1AA3 OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Ito, Y.]] | [[Category: Ito, Y.]] | ||
[[Category: Kurumizaka, H.]] | [[Category: Kurumizaka, H.]] | ||
| - | [[Category: RSGI, RIKEN | + | [[Category: RSGI, RIKEN Structural Genomics/Proteomics Initiative.]] |
[[Category: Shibata, T.]] | [[Category: Shibata, T.]] | ||
[[Category: Terada, T.]] | [[Category: Terada, T.]] | ||
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[[Category: structural genomics]] | [[Category: structural genomics]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 11:42:35 2008'' |
Revision as of 09:42, 21 February 2008
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C-TERMINAL DOMAIN OF THE E. COLI RECA, NMR, MINIMIZED AVERAGE STRUCTURE
Overview
RecA protein and its homologs catalyze homologous pairing of dsDNA and ssDNA, a critical reaction in homologous genetic recombination in various organisms from a virus, microbes to higher eukaryotes. In this reaction, RecA protein forms a nucleoprotein filament on ssDNA, which in turn binds to naked dsDNA for homology search. We suggested that the C-terminal domain of RecA protein plays a role in capturing the dsDNA. Here, we isolated the C-terminal domain as a soluble form and determined the solution structure by NMR spectroscopy. The overall folding of the NMR structure agrees with that of the corresponding part of the reported crystal structure, but a remarkable difference was found in a solvent-exposed region due to intermolecular contacts in the crystal. Then, we studied the interaction between the C-terminal domain and DNA, and found that significant chemical shift changes were induced in a specific region by titration with dsDNA. SsDNA induced a much smaller chemical shift perturbation. The difference of DNA concentrations to give the half-saturation of the chemical shift change showed a higher affinity of the C-terminal region toward dsDNA. Combined with our previous results, these provide direct evidence that the defined region in the C-terminal domain furnishes a binding surface for DNA.
About this Structure
1AA3 is a Single protein structure of sequence from Escherichia coli. Full crystallographic information is available from OCA.
Reference
An interaction between a specified surface of the C-terminal domain of RecA protein and double-stranded DNA for homologous pairing., Aihara H, Ito Y, Kurumizaka H, Terada T, Yokoyama S, Shibata T, J Mol Biol. 1997 Nov 28;274(2):213-21. PMID:9398528
Page seeded by OCA on Thu Feb 21 11:42:35 2008
Categories: Escherichia coli | Single protein | Aihara, H. | Ito, Y. | Kurumizaka, H. | RSGI, RIKEN Structural Genomics/Proteomics Initiative. | Shibata, T. | Terada, T. | Yokoyama, S. | Double-stranded dna binding domain | Riken structural genomics/proteomics initiative | Rsgi | Structural genomics
