3P9W: Crystal Structure Of An Engineered Human Autonomous Vh Domain In Complex With Vegf

Citation:
Abstract
We compared the capacity of an autonomous heavy chain variable (VH) domain (VH-B1a) to support diversity within its antigen-binding site relative to the conventional antigen-binding fragment (Fab) from which it was derived. We find that VH-B1a can tolerate significant diversity within all three complementarity-determining regions (CDRs) and also within framework 3, and thus, VH-B1a and the Fab are similar in terms of the regions of the antigen-binding site that can tolerate diversity without compromising stability. We constructed libraries of synthetic VH domains and isolated binders with moderate affinity for vascular endothelial growth factor (VEGF) from a library in which only CDR3 was randomized. One binder was subjected to affinity maturation to derive an autonomous VH domain (VH-V1a) that recognized both human and mouse VEGF with high affinity (KD=16nM or 10nM, respectively). Structural analysis revealed that VH-V1a binds to an epitope that is distinct from the epitopes of a natural VEGF receptor and six different anti-VEGF Fabs. Moreover, VH-V1a recognizes VEGF by using an unusual paratope consisting predominantly of CDR3 but with significant contributions from framework residues within the former light chain interface. These results suggest that VH-B1a and other autonomous VH domains may be useful scaffolds to support both conventional libraries with antigen-binding sites built from the three CDR loops and, also, nonconventional libraries with antigen-binding sites built from CDR3 and the former light chain interface.
PDB ID: 3P9WDownload
MMDB ID: 98787
PDB Deposition Date: 2010/10/18
Updated in MMDB: 2012/07
Experimental Method:
x-ray diffraction
Resolution: 2.41  Å
Source Organism:
Similar Structures:
Biological Unit for 3P9W: dimeric; determined by author
Molecular Components in 3P9W
Label Count Molecule
Proteins (2 molecules)
1
Vascular Endothelial Growth Factor a(Gene symbol: VEGFA)
Molecule annotation
1
Human Vegf
Molecule annotation
* Click molecule labels to explore molecular sequence information.

Citing MMDB
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