Details

Autor(en) / Beteiligte

• Boesen, Thomas P
• Soni, Bobby
• Schwartz, Thue W
• Halkier, Torben

Titel

Single-chain Vascular Endothelial Growth Factor Variant with Antagonist Activity

Ist Teil von

• The Journal of biological chemistry, 2002-10, Vol.277 (43), p.40335-40341

Ort / Verlag

United States: American Society for Biochemistry and Molecular Biology

Erscheinungsjahr

2002

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• Vascular endothelial growth factor is a specific endothelial cell mitogen that is essential for the formation of the vascular system but in the adult individual is involved in several pathological conditions, including cancer. It is a homodimeric protein that activates its receptor by binding two receptor molecules and inducing dimerization. By mixing two vascular endothelial growth factor monomers, each with different substitutions, heterodimers with only one active receptor binding site have previously been prepared. These heterodimers bind the receptor molecule but are unable to induce dimerization and activation. However, preparation of heterodimers is cumbersome, involving separate expression of different monomers, refolding the mixture, and separating heterodimers from homodimers. Here we show that a fully functional ligand can efficiently be expressed as a single protein chain containing two monomers. Single-chain vascular endothelial growth factor is functionally equivalent to the wild-type protein. By monomer-specific mutagenesis, one receptor binding site was altered. This variant competitively and specifically antagonizes the mitogenic effect of the wild-type protein on endothelial cells. The results obtained with the single-chain antagonist show the feasibility of the single-chain approach in directing alterations to single specific regions in natural homodimeric proteins that would be impossible to target in other ways.