title: Discovery of a novel vascular endothelial growth factor (VEGF) with no affinity to heparin in Gloydius tsushimaensis venom.
abstract: Strong vascular permeability enhancing activity was found only in the venom of Gloydius tsushimaensis, in Tsushima island, Japan, when examined together with the venoms of G. blomhoffii snakes in several areas of Japan and of G. ussuriensis in South Korea. The active protein purified by using Superdex 75 and Mono Q columns showed no affinity to heparin, and migrated on SDS-PAGE with molecular weights of 26 and 13 kDa under nonreducing and reducing conditions, respectively, showing that it exists as homodimer. Its N-terminal amino acid sequence was highly homologous to those of snake venom vascular endothelial growth factors (VEGFs). The sequence of this protein, named GtVF, was inferred from the one base-substituted two cDNAs (438 bp) obtained via the 3' RACE. The phylogenetic analysis suggested the presence of a new type of snake venom VEGFs including GtVF with no affinity to heparin in addition to the known three types of snake venom VEGFs with high affinity to heparin. Since the vascular permeability enhancement by GtVF was inhibited by the antibody against kinase insert domain-containing receptor (KDR), the vascular permeability enhancing activity of GtVF arises through KDR but without heparin binding.
authors.affiliation: Laboratory of Biological Chemistry, Department of Pharmaceutical Sciences, Faculty of Pharmaceutical Sciences, Sojo University, 4-22-1 Ikeda, Nishi-ku, Kumamoto 860-0082, Japan.
authors.forename: Hitomi
authors.initials: H
authors.lastname: Nakamura
authors.affiliation: Department of Applied Life Science, Faculty of Bioscience and Biotechnology, Sojo University, 4-22-1 Ikeda, Nishi-ku, Kumamoto 860-0082, Japan.
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Murakami
authors.affiliation: Department of Molecular Pathology, Faculty of Life Sciences, School of Medicine, Kumamoto University, 1-1-1 Honjo, Chuo-ku, Kumamoto 860-8556, Japan.
authors.forename: Takahisa
authors.initials: T
authors.lastname: Imamura
authors.affiliation: The Japan Snake Institute, 3318 Yabuzuka-cho, Ohta, Gunma 379-2301, Japan.
authors.forename: Michihisa
authors.initials: M
authors.lastname: Toriba
authors.affiliation: Department of Applied Life Science, Faculty of Bioscience and Biotechnology, Sojo University, 4-22-1 Ikeda, Nishi-ku, Kumamoto 860-0082, Japan.
authors.forename: Takahito
authors.initials: T
authors.lastname: Chijiwa
authors.affiliation: Department of Applied Life Science, Faculty of Bioscience and Biotechnology, Sojo University, 4-22-1 Ikeda, Nishi-ku, Kumamoto 860-0082, Japan.
authors.forename: Motonori
authors.initials: M
authors.lastname: Ohno
authors.affiliation: Laboratory of Biological Chemistry, Department of Pharmaceutical Sciences, Faculty of Pharmaceutical Sciences, Sojo University, 4-22-1 Ikeda, Nishi-ku, Kumamoto 860-0082, Japan. Electronic address: naoko@ph.sojo-u.ac.jp.
authors.forename: Naoko
authors.initials: N
authors.lastname: Oda-Ueda
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002199:Capillary Permeability / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D006168:Guinea Pigs; D006493:Heparin / Q000737:chemistry*; D007564:Japan; D008969:Molecular Sequence Data; D010802:Phylogeny; D016415:Sequence Alignment; D042461:Vascular Endothelial Growth Factor A / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D014757:Viper Venoms / Q000737:chemistry*; D017819:Viperidae
chemical_list: D042461:Vascular Endothelial Growth Factor A; D014757:Viper Venoms; D006493:Heparin
keywords: Gloydius; Heparin binding; Snake venom; VEGF; Vascular permeability enhancing activity