1ft0
CRYSTAL STRUCTURE OF TRUNCATED HUMAN RHOGDI K113A MUTANT
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OverviewOverview
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 StructureAbout this Structure
1FT0 is a Single protein structure of sequence from Homo sapiens. Full crystallographic information is available from OCA.
ReferenceReference
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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