title: NMR structure of the human oncofoetal fibronectin ED-B domain, a specific marker for angiogenesis.
abstract: BACKGROUND
  The process of angiogenesis (i.e. the formation of new blood vessels from pre-existing ones) is fundamental to physiological processes such as reproduction, development and repair, as well as to pathological conditions such as tumor progression, rheumathoid arthritis and ocular disorders. The oncofoetal ED-B domain, a specific marker of angiogenesis, consists of 91 amino acid residues that are inserted by alternative splicing into the fibronectin (FN) molecule.
  
  
  RESULTS
  The NMR structure of the ED-B domain is reported and reveals important differences from other FN type III domains. A comparison of the ED-B domain with the crystal structure of a four-domain FN fragment shows the novel features of ED-B to be located in loop regions that are buried at interdomain interfaces, and which therefore largely determine the global shape of the FN molecule. The negatively charged amino acids in this highly acidic protein are uniformly distributed over the molecular surface, with the sole exception of a solvent-exposed hydrophobic patch that represents a potential specific recognition site. Epitope mapping with 82 decapeptides that span the ED-B sequence revealed that three ED-B-specific monoclonal antibodies, which selectively target newly forming blood vessels in tumor-bearing mice, bind to adjacent regions on the ED-B surface.
  
  
  CONCLUSIONS
  The NMR structure enables the identification of a large surface area of the ED-B domain that appears to be accessible in vivo, opening up new diagnostic and therapeutic opportunities. Furthermore, the mapping of specific monoclonal antibodies to the three-dimensional structure of the ED-B domain, and their use in angiogenesis inhibition experiments, provides a basis for further investigation of the role of the ED-B domain in the formation of new blood vessels.
authors: Institut für Molekularbiologie und Biophysik, Eidgenössische Technische Hochschule Hönggerberg, CH-8093 Zürich, Switzerland.
authors: R
authors: R
authors: Fattorusso
authors: M
authors: M
authors: Pellecchia
authors: F
authors: F
authors: Viti
authors: P
authors: P
authors: Neri
authors: D
authors: D
authors: Neri
authors: K
authors: K
authors: Wüthrich
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000951:Antigens, Neoplasm / Q000737:chemistry* / Q000276:immunology / Q000502:physiology; D001665:Binding Sites; D015415:Biomarkers; D000939:Epitopes / Q000276:immunology; D005353:Fibronectins / Q000737:chemistry* / Q000276:immunology / Q000502:physiology; D017127:Glycation End Products, Advanced; D006031:Glycosylation; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008819:Mice, Nude; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009368:Neoplasm Transplantation; D009389:Neovascularization, Pathologic / Q000378:metabolism*; D011487:Protein Conformation; D020033:Protein Isoforms / Q000737:chemistry* / Q000276:immunology / Q000502:physiology; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000276:immunology / Q000502:physiology; D018243:Teratocarcinoma / Q000098:blood supply / Q000378:metabolism / Q000473:pathology; D000093844:Glycated Proteins
chemical_list: D000951:Antigens, Neoplasm; D015415:Biomarkers; D000939:Epitopes; D005353:Fibronectins; D017127:Glycation End Products, Advanced; D020033:Protein Isoforms; D011993:Recombinant Fusion Proteins; C045099:glycated fibronectin; D000093844:Glycated Proteins; C031192:oncofetal antigens