PDBsum entry 2jdf

Structural protein

PDB id: 2jdf

Contents

Protein chain

176 a.a. *

Waters ×238

* Residue conservation analysis

PDB id:

2jdf

Name:

Structural protein

Title:

Human gamma-b crystallin

Structure:

Gamma crystallin b. Chain: a. Synonym: gamma-b crystallin, gamma crystallin 1-2. Engineered: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

1.70Å R-factor: 0.196 R-free: 0.220

Authors:

H.Ebersbach,E.Fiedler,T.Scheuermann,M.Fiedler,M.T.Stubbs,C.Reimann, G.Proetzel,R.Rudolph,U.Fiedler

Key ref:

H.Ebersbach et al. (2007). Affilin-novel binding molecules based on human gamma-B-crystallin, an all beta-sheet protein. J Mol Biol, 372, 172-185. PubMed id: 17628592 DOI: 10.1016/j.jmb.2007.06.045

Date:

08-Jan-07 Release date: 24-Jul-07

Protein chain

P07316  (CRGB_HUMAN) -  Gamma-crystallin B from Homo sapiens

Seq: 175 a.a.

Struc: 176 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: E.C.?

DOI no: 10.1016/j.jmb.2007.06.045

J Mol Biol 372:172-185 (2007)

PubMed id: 17628592

Affilin-novel binding molecules based on human gamma-B-crystallin, an all beta-sheet protein.

H.Ebersbach, E.Fiedler, T.Scheuermann, M.Fiedler, M.T.Stubbs, C.Reimann, G.Proetzel, R.Rudolph, U.Fiedler.

ABSTRACT

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.

Selected figure(s)

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/].

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].

The above figures are reprinted by permission from Elsevier: J Mol Biol (2007, 372, 172-185) copyright 2007.

Figures were selected by an automated process.