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PDBsum entry 2ojr
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Protein binding
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PDB id
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2ojr
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Contents |
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* Residue conservation analysis
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DOI no:
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J Am Chem Soc
129:7114-7120
(2007)
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PubMed id:
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Double-lanthanide-binding tags for macromolecular crystallographic structure determination.
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N.R.Silvaggi,
L.J.Martin,
H.Schwalbe,
B.Imperiali,
K.N.Allen.
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ABSTRACT
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A double-lanthanide-binding tag (dLBT), a small peptide sequence engineered to
bind two lanthanide ions (e.g., Tb3+) with high affinity, was used to solve the
phase problem for the structure determination of ubiquitin by the
single-wavelength anomalous diffraction (SAD) method. Since the dLBT is
comprised exclusively of encoded amino acids, the necessity for the
incorporation of unnatural amino acids or chemical modification of the protein
as a prerequisite for X-ray structure determination is eliminated. A construct
encoding the dLBT as an N-terminal fusion with ubiquitin provides for facile
expression and purification using standard methods. Phasing of the
single-wavelength X-ray data (at 2.6 A resolution) using only the anomalous
signal from the two tightly bound Tb3+ ions in the dLBT led to clear
electron-density maps. Nearly 75% of the ubiquitin structure was built using
automated model-building software without user intervention. It is anticipated
that this technique will be broadly applicable, complementing existing
macromolecular phasing methodologies. The dLBT should be particularly useful in
cases where protein derivatization with heavy atoms proves to be problematic or
synchrotron facilities are unavailable.
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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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R.Talon,
R.Kahn,
M.A.Durá,
O.Maury,
F.M.Vellieux,
B.Franzetti,
and
E.Girard
(2011).
Using lanthanoid complexes to phase large macromolecular assemblies.
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J Synchrotron Radiat,
18,
74-78.
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C.W.am Ende,
H.Y.Meng,
M.Ye,
A.K.Pandey,
and
N.J.Zondlo
(2010).
Design of lanthanide fingers: compact lanthanide-binding metalloproteins.
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Chembiochem,
11,
1738-1747.
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G.Pompidor,
O.Maury,
J.Vicat,
and
R.Kahn
(2010).
A dipicolinate lanthanide complex for solving protein structures using anomalous diffraction.
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Acta Crystallogr D Biol Crystallogr,
66,
762-769.
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PDB codes:
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A.Thibon,
and
V.C.Pierre
(2009).
Principles of responsive lanthanide-based luminescent probes for cellular imaging.
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Anal Bioanal Chem,
394,
107-120.
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K.L.Haas,
and
K.J.Franz
(2009).
Application of metal coordination chemistry to explore and manipulate cell biology.
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Chem Rev,
109,
4921-4960.
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M.D.Hartley,
A.Larkin,
and
B.Imperiali
(2008).
Chemoenzymatic synthesis of polyprenyl phosphates.
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Bioorg Med Chem,
16,
5149-5156.
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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
codes are
shown on the right.
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Links
