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PDBsum entry 2g7b
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Transport protein
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PDB id
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2g7b
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Contents |
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* Residue conservation analysis
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J Am Chem Soc
129:6140-6148
(2007)
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PubMed id:
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Protein design: reengineering cellular retinoic acid binding protein II into a rhodopsin protein mimic.
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C.Vasileiou,
S.Vaezeslami,
R.M.Crist,
M.Rabago-Smith,
J.H.Geiger,
B.Borhan.
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ABSTRACT
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Rational redesign of the binding pocket of Cellular Retinoic Acid Binding
Protein II (CRABPII) has provided a mutant that can bind retinal as a protonated
Schiff base, mimicking the binding observed in rhodopsin. The reengineering was
accomplished through a series of choreographed manipulations to ultimately
orient the reactive species (the epsilon-amino group of Lys132 and the carbonyl
of retinal) in the proper geometry for imine formation. The guiding principle
was to achieve the appropriate Bürgi-Dunitz trajectory for the reaction to
ensue. Through crystallographic analysis of protein mutants incapable of forming
the requisite Schiff base, a highly ordered water molecule was identified as a
key culprit in orienting retinal in a nonconstructive manner. Removal of the
ordered water, along with placing reinforcing mutations to favor the desired
orientation of retinal, led to a triple mutant CRABPII protein capable of
nanomolar binding of retinal as a protonated Schiff base. The high-resolution
crystal structure of all-trans-retinal bound to the CRABPII triple mutant (1.2 A
resolution) unequivocally illustrates the imine formed between retinal and the
protein.
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Literature references that cite this PDB file's key reference
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PubMed id
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Reference
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A.N.Alexandrova,
and
W.L.Jorgensen
(2009).
Origin of the activity drop with the E50D variant of catalytic antibody 34E4 for Kemp elimination.
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J Phys Chem B,
113,
497-504.
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C.Vasileiou,
K.S.Lee,
R.M.Crist,
S.Vaezeslami,
S.M.Goins,
J.H.Geiger,
and
B.Borhan
(2009).
Dissection of the critical binding determinants of cellular retinoic acid binding protein II by mutagenesis and fluorescence binding assay.
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Proteins,
76,
281-290.
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C.Vasileiou,
W.Wang,
X.Jia,
K.S.Lee,
C.T.Watson,
J.H.Geiger,
and
B.Borhan
(2009).
Elucidating the exact role of engineered CRABPII residues for the formation of a retinal protonated Schiff base.
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Proteins,
77,
812-822.
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PDB code:
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A.N.Alexandrova,
D.Röthlisberger,
D.Baker,
and
W.L.Jorgensen
(2008).
Catalytic mechanism and performance of computationally designed enzymes for Kemp elimination.
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J Am Chem Soc,
130,
15907-15915.
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S.Vaezeslami,
X.Jia,
C.Vasileiou,
B.Borhan,
and
J.H.Geiger
(2008).
Structural analysis of site-directed mutants of cellular retinoic acid-binding protein II addresses the relationship between structural integrity and ligand binding.
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Acta Crystallogr D Biol Crystallogr,
64,
1228-1239.
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PDB codes:
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The most recent references are shown first.
Citation data come partly from CiteXplore and partly
from an automated harvesting procedure. Note that this is likely to be
only a partial list as not all journals are covered by
either method. However, we are continually building up the citation data
so more and more references will be included with time.
Where a reference describes a PDB structure, the PDB
code is
shown on the right.
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Links
