Chemical synthesis and X-ray structure of a heterochiral {D-protein antagonist plus vascular endothelial growth factor} protein complex by racemic crystallography

• DOI: 10.1073/pnas.1210483109
• Publication Date: 2012-08-28T03:18:09Z
• Authors: Kalyaneswar Mandal; Maruti Uppalapati; Dana Ault-Riché; John Kenney; Joshua Lowitz; Sachdev S. Sidhu; Stephen B.H. Kent
• Keywords: Protein ligand

Total chemical synthesis was used to prepare the mirror image ( D -protein) form of angiogenic protein vascular endothelial growth factor (VEGF-A). Phage display against -VEGF-A screen designed libraries based on a unique small scaffold in order identify high affinity ligand. Chemically synthesized - and L forms ligand showed reciprocal chiral specificity surface plasmon resonance binding experiments: The -protein bound only -VEGF-A, whereas -VEGF-A. ligand, but not inhibited natural VEGF 165 VEGFR1 receptor. Racemic crystallography determine resolution X-ray structure heterochiral complex consisting { antagonist + ofVEGF-A}. Crystallization racemic mixture these synthetic proteins appropriate stoichiometry gave more than 73 kDa containing six molecules. determined 1.6 Å. Detailed analysis interaction between VEGF-A molecule that interface comprised contact area approximately 800 Å 2 accord with our design objectives, binds same region interacts VEGFR1-domain 2.