title: Relationship between structural flexibility and function in the C-terminal region of the heparin-binding domain of VEGF165.
abstract: Vascular endothelial growth factor (VEGF) is an angiogenic protein with neurotrophic and neuroprotective effects. Previously, we reported that triamterene (Trm) inhibits VEGF-amyloid β (Aβ) interactions without affecting other biological activities of VEGF or Aβ [Jeong, K.-W., et al. (2011) Biochemistry 50, 4843-4854]. We further showed that molecular motions in the N-terminal disordered loop region of the heparin-binding domain (HBD) are important for interaction with Trm. To investigate the importance of motion at the C-terminal domain of HBD, we constructed a binding model of HBD with heparin octasaccharide (HOS) based on measurements of chemical shift changes and docking studies. Furthermore, the dynamic properties of the HBD-HOS and HBD-Trm-HOS complexes were assessed by measuring spin relaxation rates. The results showed that the HOS-binding site is composed of two basic clusters consisting of side chains of residues R13, R14, and K15 and residues K30, R35, and R49. When HOS binds, values for the heteronuclear nuclear Overhauser effect near HOS-binding sites increased dramatically. CPMG (Carr-Purcell-Meiboom-Gill sequence) experiments as well as an R2 relaxation experiment were undertaken to understand millisecond time-scale motions in HBD. There is large relaxation dispersion of residues at Trm- and HOS-binding sites in free HBD. C-Terminal residues such as S34, C48, and D51 near the HOS-binding sites continued to exhibit slow conformational motions in the HBD-Trm complex, while those slow motions disappeared in the bound conformation of HBD with HOS. Collectively, our results demonstrate that the inherent structural flexibilities of the C-terminal region of the HBD are important in the heparin binding process and that Trm does not inhibit VEGF-heparin interactions necessary for the biological activities of VEGF.
authors.affiliation: Department of Bioscience and Biotechnology and Bio/Molecular Informatics Center, Institute of KU Biotechnology, Konkuk University , Seoul 143-701, Korea.
authors.forename: Ki-Woong
authors.initials: KW
authors.lastname: Jeong
authors.forename: Min-Cheol
authors.initials: MC
authors.lastname: Jeong
authors.forename: Bonghwan
authors.initials: B
authors.lastname: Jin
authors.forename: Yangmee
authors.initials: Y
authors.lastname: Kim
mesh_terms: D001665:Binding Sites; D006493:Heparin / Q000737:chemistry*; D006801:Humans; D006860:Hydrogen Bonding; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013499:Surface Properties; D042461:Vascular Endothelial Growth Factor A / Q000737:chemistry*
chemical_list: C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D006493:Heparin