1ft0
From Proteopedia
(New page: 200px<br /> <applet load="1ft0" size="450" color="white" frame="true" align="right" spinBox="true" caption="1ft0, resolution 2.6Å" /> '''CRYSTAL STRUCTURE OF...) |
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| - | [[Image:1ft0.gif|left|200px]]<br /> | + | [[Image:1ft0.gif|left|200px]]<br /><applet load="1ft0" size="350" color="white" frame="true" align="right" spinBox="true" |
| - | <applet load="1ft0" size=" | + | |
caption="1ft0, resolution 2.6Å" /> | caption="1ft0, resolution 2.6Å" /> | ||
'''CRYSTAL STRUCTURE OF TRUNCATED HUMAN RHOGDI K113A MUTANT'''<br /> | '''CRYSTAL STRUCTURE OF TRUNCATED HUMAN RHOGDI K113A MUTANT'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Crystallization is a unique process that occurs at the expense of entropy, including the conformational entropy of surface residues, which become | + | Crystallization is a unique process that occurs at the expense of entropy, including the conformational entropy of surface residues, which become ordered in crystal lattices during formation of crystal contacts. It could therefore be argued that epitopes free of amino acids with high conformational entropy are more thermodynamically favorable for crystal formation. For a protein recalcitrant to crystallization, mutation of such surface amino acids to residues with no conformational entropy might lead to enhancement of crystallization. This paper reports the results of experiments with an important cytosolic regulator of GTPases, human RhoGDI, in which lysine residues were systematically mutated to alanines. Single and multiple mutations were introduced into two different variants of RhoGDI, NDelta23 and NDelta66, in which the first 23 and 66 residues, respectively, were removed by recombinant methods. In total, 13 single and multiple mutants were prepared and assessed for crystallization and all were shown to crystallize using the Hampton Research Crystal Screens I and II, in contrast to wild-type NDelta23 and NDelta66 RhoGDI which did not crystallize. Four crystal structures were solved (the triple mutants NDelta23:K135,138,141A and NDelta66:K135,138,141A, and two single mutants NDelta66:K113A and NDelta66:K141A) and in three cases the crystal contacts of the new lattices were found precisely at the sites of mutations. These results support the notion that it is, in principle, possible to rationally design mutations which systematically enhance proteins' ability to crystallize. |
==About this Structure== | ==About this Structure== | ||
| - | 1FT0 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens]. Full crystallographic information is available from [http:// | + | 1FT0 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1FT0 OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Homo sapiens]] | [[Category: Homo sapiens]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
| - | [[Category: Derewenda, Z | + | [[Category: Derewenda, Z S.]] |
| - | [[Category: Garrard, S | + | [[Category: Garrard, S M.]] |
| - | [[Category: Longenecker, K | + | [[Category: Longenecker, K L.]] |
| - | [[Category: Sheffield, P | + | [[Category: Sheffield, P J.]] |
[[Category: beta sandwich motif]] | [[Category: beta sandwich motif]] | ||
[[Category: gdp-dissociation inhibitor of rho gtpases]] | [[Category: gdp-dissociation inhibitor of rho gtpases]] | ||
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[[Category: isoprenyl-binding domain]] | [[Category: isoprenyl-binding domain]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 12:42:18 2008'' |
Revision as of 10:42, 21 February 2008
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CRYSTAL STRUCTURE OF TRUNCATED HUMAN RHOGDI K113A MUTANT
Overview
Crystallization is a unique process that occurs at the expense of entropy, including the conformational entropy of surface residues, which become ordered in crystal lattices during formation of crystal contacts. It could therefore be argued that epitopes free of amino acids with high conformational entropy are more thermodynamically favorable for crystal formation. For a protein recalcitrant to crystallization, mutation of such surface amino acids to residues with no conformational entropy might lead to enhancement of crystallization. This paper reports the results of experiments with an important cytosolic regulator of GTPases, human RhoGDI, in which lysine residues were systematically mutated to alanines. Single and multiple mutations were introduced into two different variants of RhoGDI, NDelta23 and NDelta66, in which the first 23 and 66 residues, respectively, were removed by recombinant methods. In total, 13 single and multiple mutants were prepared and assessed for crystallization and all were shown to crystallize using the Hampton Research Crystal Screens I and II, in contrast to wild-type NDelta23 and NDelta66 RhoGDI which did not crystallize. Four crystal structures were solved (the triple mutants NDelta23:K135,138,141A and NDelta66:K135,138,141A, and two single mutants NDelta66:K113A and NDelta66:K141A) and in three cases the crystal contacts of the new lattices were found precisely at the sites of mutations. These results support the notion that it is, in principle, possible to rationally design mutations which systematically enhance proteins' ability to crystallize.
About this Structure
1FT0 is a Single protein structure of sequence from Homo sapiens. Full crystallographic information is available from OCA.
Reference
Protein crystallization by rational mutagenesis of surface residues: Lys to Ala mutations promote crystallization of RhoGDI., Longenecker KL, Garrard SM, Sheffield PJ, Derewenda ZS, Acta Crystallogr D Biol Crystallogr. 2001 May;57(Pt 5):679-88. Epub 2001, Apr 24. PMID:11320308
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