1sj5
From Proteopedia
(New page: 200px<br /><applet load="1sj5" size="450" color="white" frame="true" align="right" spinBox="true" caption="1sj5, resolution 2.80Å" /> '''Crystal structure of...) |
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| - | [[Image:1sj5.gif|left|200px]]<br /><applet load="1sj5" size=" | + | [[Image:1sj5.gif|left|200px]]<br /><applet load="1sj5" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1sj5, resolution 2.80Å" /> | caption="1sj5, resolution 2.80Å" /> | ||
'''Crystal structure of a C38A mutant of a protein with unknown function from DUF151 family (TM0160) from Thermotoga maritima at 2.8 A resolution'''<br /> | '''Crystal structure of a C38A mutant of a protein with unknown function from DUF151 family (TM0160) from Thermotoga maritima at 2.8 A resolution'''<br /> | ||
==Overview== | ==Overview== | ||
| - | The structure of two Thermotoga maritima proteins, a conserved | + | The structure of two Thermotoga maritima proteins, a conserved hypothetical protein (TM0160) and a transcriptional regulator (TM1171), have now been determined at 1.9 A and 2.3 A resolution, respectively, as part of a large-scale structural genomics project. Our first efforts to crystallize full-length versions of these targets were unsuccessful. However, analysis of the recombinant purified proteins using the technique of enhanced amide hydrogen/deuterium exchange mass spectroscopy (DXMS) revealed substantial regions of rapid amide deuterium hydrogen exchange, consistent with flexible regions of the structures. Based on these exchange data, truncations were designed to selectively remove the disordered C-terminal regions, and the resulting daughter proteins showed greatly enhanced crystallizability. Comparative DXMS analysis of full-length protein versus truncated forms demonstrated complete and exact preservation of the exchange rate profiles in the retained sequence, indicative of conservation of the native folded structure. This study presents the first structures produced with the aid of the DXMS method for salvaging intractable crystallization targets. The structure of TM0160 represents a new fold and highlights the use of this approach where any prior structural knowledge is absent. The structure of TM1171 represents an example where the lack of a substrate/cofactor may impair crystallization. The details of both structures are presented and discussed. |
==About this Structure== | ==About this Structure== | ||
| - | 1SJ5 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Thermotoga_maritima Thermotoga maritima]. Full crystallographic information is available from [http:// | + | 1SJ5 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Thermotoga_maritima Thermotoga maritima]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1SJ5 OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Thermotoga maritima]] | [[Category: Thermotoga maritima]] | ||
| - | [[Category: JCSG, Joint | + | [[Category: JCSG, Joint Center for Structural Genomics.]] |
| - | [[Category: Jr., V | + | [[Category: Jr., V L.Woods.]] |
| - | [[Category: Klock, H | + | [[Category: Klock, H E.]] |
| - | [[Category: Lesley, S | + | [[Category: Lesley, S A.]] |
[[Category: Panatazatos, D.]] | [[Category: Panatazatos, D.]] | ||
[[Category: Spraggon, G.]] | [[Category: Spraggon, G.]] | ||
| - | [[Category: Wilson, I | + | [[Category: Wilson, I A.]] |
[[Category: dxms]] | [[Category: dxms]] | ||
[[Category: hypothetical protein]] | [[Category: hypothetical protein]] | ||
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[[Category: tm0160]] | [[Category: tm0160]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 15:02:03 2008'' |
Revision as of 13:02, 21 February 2008
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Crystal structure of a C38A mutant of a protein with unknown function from DUF151 family (TM0160) from Thermotoga maritima at 2.8 A resolution
Overview
The structure of two Thermotoga maritima proteins, a conserved hypothetical protein (TM0160) and a transcriptional regulator (TM1171), have now been determined at 1.9 A and 2.3 A resolution, respectively, as part of a large-scale structural genomics project. Our first efforts to crystallize full-length versions of these targets were unsuccessful. However, analysis of the recombinant purified proteins using the technique of enhanced amide hydrogen/deuterium exchange mass spectroscopy (DXMS) revealed substantial regions of rapid amide deuterium hydrogen exchange, consistent with flexible regions of the structures. Based on these exchange data, truncations were designed to selectively remove the disordered C-terminal regions, and the resulting daughter proteins showed greatly enhanced crystallizability. Comparative DXMS analysis of full-length protein versus truncated forms demonstrated complete and exact preservation of the exchange rate profiles in the retained sequence, indicative of conservation of the native folded structure. This study presents the first structures produced with the aid of the DXMS method for salvaging intractable crystallization targets. The structure of TM0160 represents a new fold and highlights the use of this approach where any prior structural knowledge is absent. The structure of TM1171 represents an example where the lack of a substrate/cofactor may impair crystallization. The details of both structures are presented and discussed.
About this Structure
1SJ5 is a Single protein structure of sequence from Thermotoga maritima. Full crystallographic information is available from OCA.
Reference
On the use of DXMS to produce more crystallizable proteins: structures of the T. maritima proteins TM0160 and TM1171., Spraggon G, Pantazatos D, Klock HE, Wilson IA, Woods VL Jr, Lesley SA, Protein Sci. 2004 Dec;13(12):3187-99. PMID:15557262
Page seeded by OCA on Thu Feb 21 15:02:03 2008
Categories: Single protein | Thermotoga maritima | JCSG, Joint Center for Structural Genomics. | Jr., V L.Woods. | Klock, H E. | Lesley, S A. | Panatazatos, D. | Spraggon, G. | Wilson, I A. | Dxms | Hypothetical protein | Jcsg | Joint center for structural genomics | Protein structure initiative | Psi | Structural genomics | Tm0160
