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PDBsum entry 2jhs
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References listed in PDB file
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Key reference
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Title
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Protein crystallization by surface entropy reduction: optimization of the ser strategy.
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Authors
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D.R.Cooper,
T.Boczek,
K.Grelewska,
M.Pinkowska,
M.Sikorska,
M.Zawadzki,
Z.Derewenda.
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Ref.
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Acta Crystallogr D Biol Crystallogr, 2007,
63,
636-645.
[DOI no: ]
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PubMed id
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Abstract
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A strategy of rationally engineering protein surfaces with the aim of obtaining
mutants that are distinctly more susceptible to crystallization than the
wild-type protein has previously been suggested. The strategy relies on
replacing small clusters of two to three surface residues characterized by high
conformational entropy with alanines. This surface entropy reduction (or SER)
method has proven to be an effective salvage pathway for proteins that are
difficult to crystallize. Here, a systematic comparison of the efficacy of using
Ala, His, Ser, Thr and Tyr to replace high-entropy residues is reported. A total
of 40 mutants were generated and screened using two different procedures. The
results reaffirm that alanine is a particularly good choice for a replacement
residue and identify tyrosines and threonines as additional candidates that have
considerable potential to mediate crystal contacts. The propensity of these
mutants to form crystals in alternative screens in which the normal
crystallization reservoir solutions were replaced with 1.5 M NaCl was also
examined. The results were impressive: more than half of the mutants yielded a
larger number of crystals with salt as the reservoir solution. This method
greatly increased the variety of conditions that yielded crystals. Taken
together, these results suggest a powerful crystallization strategy that
combines surface engineering with efficient screening using standard and
alternate reservoir solutions.
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Figure 1.
Figure 1 The distribution of the mutations used in this study.
The mutation clusters were designated by a single letter A-I.
The mutations corresponding to each cluster are shown as magenta
spheres on the ribbon diagram. A cartoon representation of
RhoGDI is shown for reference.
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Figure 4.
Figure 4 Histogram of hits in the salt screens.
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The above figures are
reprinted
by permission from the IUCr:
Acta Crystallogr D Biol Crystallogr
(2007,
63,
636-645)
copyright 2007.
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