1vjl: Difference between revisions

Revision as of 16:35, 21 February 2008

CRYSTAL STRUCTURE OF PREDICTED PROTEIN RELATED TO WOUND INDUCIVE PROTEINS IN PLANTS (TM0160) FROM THERMOTOGA MARITIMA AT 1.90 A RESOLUTION

OverviewOverview

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 StructureAbout this Structure

1VJL is a Single protein structure of sequence from Thermotoga maritima with and as ligands. This structure supersedes the now removed PDB entry 1O5Y. Full crystallographic information is available from OCA.

ReferenceReference

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 15:35:57 2008

Proteopedia Page Contributors and Editors (what is this?)Proteopedia Page Contributors and Editors (what is this?)

OCA

@@ Line 1: / Line 1: @@
-[[Image:1vjl.gif|left|200px]]<br /><applet load="1vjl" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:1vjl.gif|left|200px]]<br /><applet load="1vjl" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="1vjl, resolution 1.90&Aring;" />
 '''CRYSTAL STRUCTURE OF PREDICTED PROTEIN RELATED TO WOUND INDUCIVE PROTEINS IN PLANTS (TM0160) FROM THERMOTOGA MARITIMA AT 1.90 A RESOLUTION'''<br />
 ==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.
+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==
-VJL is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Thermotoga_maritima Thermotoga maritima] with CL and UNL as [http://en.wikipedia.org/wiki/ligands ligands]. This structure superseeds the now removed PDB entry 1O5Y. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1VJL OCA].
+VJL is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Thermotoga_maritima Thermotoga maritima] with <scene name='pdbligand=CL:'>CL</scene> and <scene name='pdbligand=UNL:'>UNL</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. This structure supersedes the now removed PDB entry 1O5Y. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1VJL OCA].
 ==Reference==
@@ Line 13: / Line 13: @@
 [[Category: Single protein]]
 [[Category: Thermotoga maritima]]
-[[Category: Genomics, Joint.Center.for.Structural.]]
+[[Category: Genomics, Joint Center for Structural.]]
-[[Category: JCSG, Joint.Center.for.Structural.Genomics.]]
+[[Category: JCSG, Joint Center for Structural Genomics.]]
 [[Category: CL]]
 [[Category: UNL]]
@@ Line 25: / Line 25: @@
 [[Category: tm0160]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Sat Nov 24 22:34:53 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 15:35:57 2008''