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PDBsum entry 2jdf
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Structural protein
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PDB id
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2jdf
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References listed in PDB file
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Key reference
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Title
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Affilin-Novel binding molecules based on human gamma-B-Crystallin, An all beta-Sheet protein.
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Authors
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H.Ebersbach,
E.Fiedler,
T.Scheuermann,
M.Fiedler,
M.T.Stubbs,
C.Reimann,
G.Proetzel,
R.Rudolph,
U.Fiedler.
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Ref.
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J Mol Biol, 2007,
372,
172-185.
[DOI no: ]
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PubMed id
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Abstract
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The concept of novel binding proteins as an alternative to antibodies has
undergone rapid development and is now ready for practical use in a wide range
of applications. Alternative binding proteins, based on suitable scaffolds with
desirable properties, are selected from combinatorial libraries in vitro. Here,
we describe an approach using a beta-sheet of human gamma-B-crystallin to
generate a universal binding site through randomization of eight solvent-exposed
amino acid residues selected according to structural and sequence analyses.
Specific variants, so-called Affilin, have been isolated from a phage display
library against a variety of targets that differ considerably in size and
structure. The isolated Affilin variants can be produced in Escherichia coli as
soluble proteins and have a high level of thermodynamic stability. The crystal
structures of the human wild-type gamma-B-crystallin and a selected Affilin
variant have been determined to 1.7 A and 2.0 A resolution, respectively.
Comparison of the two molecules indicates that the human gamma-B-crystallin
tolerates amino acid exchanges with no major structural change. We conclude that
the intrinsically stable and easily expressed gamma-B-crystallin provides a
suitable framework for the generation of novel binding molecules.
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Figure 1.
Figure 1. A representation of bovine γ-B-crystallin used to
model the human γ-B-crystallin scaffold, showing the C^α trace
in cartoon representation and the accessible surface area. The
eight selected amino acid residues (positions 2, 4, 6, 15, 17,
19, 36, and 38) used for library construction are highlighted in
red. This Figure was generated using pdb entry 1AMM and the
program PyMol [http://pymol.sourceforge.net/].
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Figure 9.
Figure 9. Superposition of all C^α backbones (right) and of
the N-terminal domain (left) of the crystal structure of human
wild-type γ-B-crystallin (red) and of the IgG-Fc binding
Affilin molecule SPC-1-G3 (blue). The superposition was
performed using the program PyMol with rms values of 0.284
Å (N-terminal domain) [http://pymol.sourceforge.net].
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The above figures are
reprinted
by permission from Elsevier:
J Mol Biol
(2007,
372,
172-185)
copyright 2007.
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