1wvl
From Proteopedia
(New page: 200px<br /><applet load="1wvl" size="450" color="white" frame="true" align="right" spinBox="true" caption="1wvl, resolution 2.6Å" /> '''Crystal Structure of ...) |
|||
| Line 1: | Line 1: | ||
| - | [[Image:1wvl.gif|left|200px]]<br /><applet load="1wvl" size=" | + | [[Image:1wvl.gif|left|200px]]<br /><applet load="1wvl" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1wvl, resolution 2.6Å" /> | caption="1wvl, resolution 2.6Å" /> | ||
'''Crystal Structure of Multimeric DNA-binding Protein Sac7d-GCN4 with DNA decamer'''<br /> | '''Crystal Structure of Multimeric DNA-binding Protein Sac7d-GCN4 with DNA decamer'''<br /> | ||
==Overview== | ==Overview== | ||
| - | The protein Sac7d belongs to a class of small chromosomal proteins from | + | The protein Sac7d belongs to a class of small chromosomal proteins from the hyperthermophilic archaeon Sulfolobus acidocaldarius. Sac7d is extremely stable to heat, acid, and chemical agents. This protein is a monomer and it binds DNA without any particular sequence preference, while inducing a sharp kink in the DNA. By appending a leucine-zipper-like helical peptide derived from the yeast transcriptional activator GCN4 to the C-terminal end of Sac7d, the modified monomers (denoted S7dLZ) are expected to interact with each other via hydrophobic force to form a parallel dimer. The recombinant S7dLZ was expressed in Escherichia coli and purified by heating and ion-exchange chromatography. The formation of dimer was detected by gel-filtration chromatography and chemical cross-link. The results of surface plasmon resonance and circular dichroism experiments showed that the DNA-binding capacity was retained. Furthermore, X-ray diffraction analysis of single crystals of S7dLZ in complex with DNA decamer CCTATATAGG showed that the leucine-zipper segments of S7dLZ were associated into an antiparallel four-helix bundle. There are two DNA fragments bound to each S7dLZ tetramer in the crystal. This model works as a successful template that endows protein a new function without losing original properties. |
==About this Structure== | ==About this Structure== | ||
| - | 1WVL is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Sulfolobus_acidocaldarius Sulfolobus acidocaldarius]. Full crystallographic information is available from [http:// | + | 1WVL is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Sulfolobus_acidocaldarius Sulfolobus acidocaldarius]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1WVL OCA]. |
==Reference== | ==Reference== | ||
| Line 13: | Line 13: | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Sulfolobus acidocaldarius]] | [[Category: Sulfolobus acidocaldarius]] | ||
| - | [[Category: Wang, A | + | [[Category: Wang, A H.]] |
| - | [[Category: Wu, S | + | [[Category: Wu, S W.]] |
[[Category: leucine zipper]] | [[Category: leucine zipper]] | ||
[[Category: protein engineering]] | [[Category: protein engineering]] | ||
| Line 21: | Line 21: | ||
[[Category: x-ray crystallography]] | [[Category: x-ray crystallography]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 15:48:34 2008'' |
Revision as of 13:48, 21 February 2008
|
Crystal Structure of Multimeric DNA-binding Protein Sac7d-GCN4 with DNA decamer
Overview
The protein Sac7d belongs to a class of small chromosomal proteins from the hyperthermophilic archaeon Sulfolobus acidocaldarius. Sac7d is extremely stable to heat, acid, and chemical agents. This protein is a monomer and it binds DNA without any particular sequence preference, while inducing a sharp kink in the DNA. By appending a leucine-zipper-like helical peptide derived from the yeast transcriptional activator GCN4 to the C-terminal end of Sac7d, the modified monomers (denoted S7dLZ) are expected to interact with each other via hydrophobic force to form a parallel dimer. The recombinant S7dLZ was expressed in Escherichia coli and purified by heating and ion-exchange chromatography. The formation of dimer was detected by gel-filtration chromatography and chemical cross-link. The results of surface plasmon resonance and circular dichroism experiments showed that the DNA-binding capacity was retained. Furthermore, X-ray diffraction analysis of single crystals of S7dLZ in complex with DNA decamer CCTATATAGG showed that the leucine-zipper segments of S7dLZ were associated into an antiparallel four-helix bundle. There are two DNA fragments bound to each S7dLZ tetramer in the crystal. This model works as a successful template that endows protein a new function without losing original properties.
About this Structure
1WVL is a Single protein structure of sequence from Sulfolobus acidocaldarius. Full crystallographic information is available from OCA.
Reference
Design and characterization of a multimeric DNA binding protein using Sac7d and GCN4 as templates., Wu SW, Ko TP, Chou CC, Wang AH, Proteins. 2005 Sep 1;60(4):617-28. PMID:16028219
Page seeded by OCA on Thu Feb 21 15:48:34 2008
