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PDBsum entry 1qvy
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Signaling protein inhibitor
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PDB id
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1qvy
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Contents |
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* Residue conservation analysis
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References listed in PDB file
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Key reference
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Title
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The impact of lys-->Arg surface mutations on the crystallization of the globular domain of rhogdi.
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Authors
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J.Czepas,
Y.Devedjiev,
D.Krowarsch,
U.Derewenda,
J.Otlewski,
Z.S.Derewenda.
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Ref.
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Acta Crystallogr D Biol Crystallogr, 2004,
60,
275-280.
[DOI no: ]
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PubMed id
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Abstract
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The potential of rational surface mutagenesis for enhanced protein
crystallization is being probed in an ongoing effort. In previous work, it was
hypothesized that residues with high conformational entropy such as Glu and Lys
are suitable targets for surface mutagenesis, as they are rarely incorporated in
crystal contacts or protein-protein interfaces. Previous experiments using
Lys-->Ala, Glu-->Ala and Glu-->Asp mutants confirmed that mutated proteins were
more likely to crystallize. In the present paper, the usefulness of Lys-->Arg
mutations is studied. Several mutations of the globular domain of human RhoGDI
were generated, including the single mutants K105R, K113R, K127R, K138R and
K141R, the double mutants K(98,99)R and K(199,200)R and the triple mutants
K(98,99,105)R and K(135,138,141)R. It is shown that Lys-->Arg mutants are more
likely to crystallize than the wild-type protein, although not as likely as
Lys-->Ala mutants. Out of the nine mutants tested, five produced diffracting
crystals, including the K(199,200)R double mutant, which crystallized in a new
space group and exceeded by approximately 1.0 A the resolution of the
diffraction of the wild-type crystal. Major crystal contacts in the new lattice
were created by the mutated epitope.
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Figure 3.
Figure 3 Electron-density map in the region of the lattice
contact contoured at the 1  level,
involving mutated Arg199, its symmetry-related pair Arg199 sym
as well as the charge constellation including the negatively
charged residues Asp204, Asp204 sym and two solvent sulfates.
Additional hydrogen bonds involving Arg200, Ser174 and their
symmetry-related partners contribute to stability of the lattice
contact. The figure was prepared with BOBSCRIPT (Esnouf,
1997[Esnouf, R. M. (1997). J. Mol. Graph. 15, 132-143.]).
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The above figure is
reprinted
by permission from the IUCr:
Acta Crystallogr D Biol Crystallogr
(2004,
60,
275-280)
copyright 2004.
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