• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to project page

1995 Fiscal Year Final Research Report Summary

Accelerated Evolution of Venom Gland Isozymes

Research Project

Project/Area Number 07044204
Research Category

Grant-in-Aid for international Scientific Research

Allocation TypeSingle-year Grants
SectionJoint Research
Research InstitutionKYUSHU UNIVERSITY

Principal Investigator

OHNO Motonori  Department of Chemistry, Faculty of Science, Kyushu University, Professor, 理学部, 教授 (30038434)

Co-Investigator(Kenkyū-buntansha) OGAWA Tomohisa  Department of Chemistry, Faculty of Science, Kyushu University, 理学部, 助手 (80240901)
SHIMOHIGASHI Yasuyuki  Department of Chemistry, Faculty of Science, Kyushu University, 理学部, 助教授 (00211293)
MENEZ Andre  Department d'ingenierie et d'Etueles des Proteines C.E.Saclay, 所長
Project Period (FY) 1995
Keywordssnake venom / molecular evolution / phospholipase A2 / neurotoxin / accelerated evolution / adaptive / cDNA / isozyme
Research Abstract

Snake venom contains a variety of physiological active enzymes which act on mammalian target molecules with high selectivety. Snakes know everything about mammalian (human) homeostasis. One of the most intriguing questions is how snake toxins have evolved to acquire such functions specific to mammalian constitutes. Recently, we discovered that crotalidae snake phospholipase A_2 (PLA_2) isozyme genes have evolved via accelerated evolution only in the mature protein-coding region to gain diverse physiological functions.
A major theme in this project is to prove the universality of this hypothesis for other snake toxins and species to elucidate the adaptive evolutionary mechanism.
First, we prepared cDNA libraries from Naja naja kaouthia and Dendroaspis angnsticeps (green mamba) venom gland. We obtained 50 positive clones encoding group I PLA_2 isozyme cDNAs from Naja naja kaouthia venom gland. Two cDNA clones were seqenced. Their nucleotide sequences encoded new PLA_2 isozymes with four amino acid differences to each other.
From D.angnsticeps venom gland cDNA library, four new cDNAs encoding "theree finger" shaped isotoxins, which also exhibit various diverse fanctions, were cloned and sequenced. The comparison of the nucleotide sequences of the "three finger" shaped isotoxin cDANs revealed that the mature protein-coding region is much more variable than the noncoding region and that the numbers of nucleotide substitutions per nosnsynonymous site (K_A) are close to or larger than the numbers of nucleotide substitutions per synonymous site (K_S) in the mature protein-coding region. These observations suggest that these isotoxins have also evolved via Darwinian-type accelerated evolution. It is apparent that accelerated eolution is universal in snake venom isozyme or isoprotein families.

  • Research Products

    (50 results)

All Other

All Publications (50 results)

  • [Publications] K.Nakashima,I.Nobuhisa,M.Deshimaru,T.Ogawa,Y.Shimohigashi,Y.Fukumaki,M.Hattori,Y.Sakaki,S.Hattori&M.Ohno: "Structures of Genes Fncoding TATABox-Binding Proteins from Trimeresurus gramineus and T.flavoviridis Snakes." Gene. 152. 209-213 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A.Suzuki,E.Matsueda,T.Yamane,T.Ashida,H.Kihara&M.Ohno: "Crystal Strueture Analysis of Phospholipase A_2 from Trimeresurus flavoviridis (Habu Snake) Venom at 1.5 A Resolution" Journal of Biochemistry. 117. 730-740 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Ogawa,K.Nakashima,N.Oda,Y.Shimohigashi,M.Ohno,S.Hattori,M.Hattori,Y.Sakaki&H.Kihara: "Trimeresurus flavoviridis Venom Phospholipase A_2 Genes Have Evolved via Accelerated Substitutions." Journal of Molecular Recognition. 8. 40-46 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Shimohigashi,T.Nose,M.Ohno,Y.Ogino&T.Costa: "Human Thrombine Receptors Are Insensitive Thrombin-Like Snake Venom." Biochemistry and Molecular Biology Int.35. 415-421 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Nakashima,I.Nobuhisa,M.Deshimaru,M.Nakai,T.Ogawa,Y.Shimohigashi,Y.Fukumaki,M.Hattori,Y.Sakaki,S.Hattori&M.Ohno: "Accelerated Evolution in the Protein-Coding Regions Is Universal in Crotalinae Snake Venom Gland Phospholipase A_2" The Proceedings of National Academy of Sciences USA. 92. 5605-5609 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Nakai,K.Nakashima,T.Ogawa,Y.Shimohigashi,S.Hattori,C.C.Chang&M.Ohno: "Purification and Primary Structure of a Myotoxic Lysine-49-Phospholipase A_2 with Low Lipolytic Activity from Trimeresurus gramineus Venom." Toxicon. 33. 1469-1478 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Shimohigashi,A.Tani,H.Matsumoto,K.Nakashima,Y.Yamaguchi,N.Oda,Y.Takano,H.Kamiya,J.Kishino,H.Arita&M.Ohno: "Lysine-49-Phospholipase A_2 from Trimeresurus flavoviridis Venom Are Membrane-Acting Enzymes" Journal of Biochemistry. 118. 1037-1044 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Ogawa,H.Onoue,K.Nakagawa,S.Nomura,K.Sueishi,S.Hattori,H.Kihra&M.Ohno: "Localization and Expression of Phospholipase A_2 in Trimeresurus flavoviridis (Habu Snake) Venom Gland." Taxicon. 33. 1645-1652 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Ogawa,M.Kitajima,K.Nakashima,Y.Sakaki&M.Ohno: "Molecular Evolution of Group II Phospholipase A_2" Journal of Molecular Evolution. 41. 867-877 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] S.Nakamura,M.Nakai,K.Nakashima,T.Ogawa,Y.Shimohigashi,H.Kihara,T.Yamane,T.Ashida&M.Ohno: "Roles of Lysine-69 in Dimerization and Activity of Trimeresurus flavoviridis Venom Asparate-49-Phospholipase A_2." Journal of Molecular Recognition. (in press). (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] I.Nobuhisa,K.Nakashima,M.Deshimaru,T.Ogawa,Y.Shimahigashi,Y.Fukumaki,Y.Sakaki,S.Hattori,H.Kihara&M.Ohno: "Accelerated Evolution of Trimeresurus Okinavensis Venom Gland Phospholipase A_2 Isozymes Genes." Gene. (in press). (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Ogawa,K.Nakashima,I.Nobuhisa,M.Deshimaru,Y.Shimohigashi,Y.Fukumaki,Y.Sakaki,S.Hattori&M.Ohno: "Accelerated Evolution of Snake Venom Phospholipase A_2 Isozymes for Acquisition of Diverse Physiological Functions." Toxicon. (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] B.Maillere,G.Mourier,J.Cotton,M.Herve,S.Leroy&A.Menez: "Probing immunogenicity of a T cell epitope by Lalanine and D-amino acid scanning." Mol Immunol. 32. 1073-1080 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Dauplais,B.Gilquin,L.D.Possani,G.Gurrola-Briones,C.Roumestand&A.Menez: "Determination of the three-dimensional solution structure of noxiustoxin : analysis of structural difference with related short-chain scorpion toxins" Biochemistry. 34. 16563-16573 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] P.Cuniasse,A.Thomas,J.C.Smith,H.L.Thanh,M.Leonetti&A.Menez: "Structural basis of antibody cross-reactivity : soulution conformation of an immunogenic peptide fragment containing both T and B epitopes." Biochemistry. 34. 12782-12789 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] B.Maillere,G.Mourier,M.Herve,J.Cotton,S.Leroy&A.Menez: "Immunogenicity of a disulphide-containing neurotoxin : presentation to T-cells requires a reduction step." Toxicon. 33. 475-482 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A.L.Harvey,H.Vatanpour,E.G.Rowan,S.Pinkasfeld,C.Vita,A.Menez&M.F.Martin-Eauclaire: "Structure-activity studies on scorpion toxins that block potassium channels." Toxicon. 33. 425-436 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] R.Genet,P.H.Benetti,A.Hammadi&A.Menez: "L-tryptophan 2',3'-oxidase from Chromobacterium violaceum. Substrate specificity and mechanistic implications." J Biol Chem. 270. 23540-23545 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] C.Vita,C.Roumestand,F.Toma&A.Menez: "Scorpion toxins as natural scaffolds for protein engineering." Proc Natl Acad Sci USA. 92. 6404-6408 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Leonetti,J.Cotton,S.Leroy,G.Mourier&A.Menez: "Immunogenicity of T epitope-containing cyclic peptides. Increasing neutalizing antibody responses by introducing fine chemical changes." J Immunol. 155. 210-218 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Dauplais,J.M.Neumann,S.Pinkasfeld,A.Menez&C.Roumestand: "An NMR study of the interraction of cardiotoxin gamma from Naja nigricollis with perdeuterated dodecylphosphocholine micelles." Eur J Biochem. 230. 213-220 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A.Carrier,F.Ducancel,N.B.Settiawan,L.Cattolico,B.Maillere,M.Leonetti,P.Drevet,A.Menez&J.C.Boulain: "Recombinant antibody-alkakine phosphatase conjugates foe diagnosis of human IgGs : application to anti-HBsAg detection." J Immunol Methods. 181. 177-186 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] O.Tremeau,C.Lemaire,P.Drevet,S.Pinkasfeld,F.Ducancel,J.C.Boulain&A.Menez: "Genetic engineering of snake toxins. The functional site of Erabutoxin a,as delineated by site-directed mutagenesis,includes variant residues." J Biol Chem. 270. 9362-9369 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] D.Servent,A.Menez&P.Kessler: "Site-directed disulfide reduction using an affinity reagent : application on the nicotinic acetylcholine receptor." FEBS Lett.360. 261-265 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] I.Segalas,C.Roumestand,S.Zinn-Justin,B.Gilquin,R.Menez,A.Menez&F.Toma20GB25:Solution structure of a green mamba toxin that activates muscarinic acetycholine receptors,as studied by nuclear magnetic resonance and molecular modeling.: Biochemistry. 34. 1248-1260 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Nakashima, I.Nobuhisa, M.Deshimaru, T.Ogawa, Y.Shimohigashi, Y.Fukumaki, M.Hattori, Y.Sakaki, S.Hattori & M.Ohno: "Structures of Genes Encoding TATA Box-Binding Proteins from Trimeresurus gramineus and T.flavoviridis Snakes." Gene. 152. 209-213 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] A.Suzuki, E.Matsueda, T.Yamane, T.Ashida, H.Kihara & M.Ohno: "Crystal Structure Analysis of Phospholipase A_2 from Trimeresurus flavoviridis (Habu Snake) Venom at 1.5@90 Resoltion." Journal of Biochemistry. 117. 730-740 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Ogawa, K.Nakashima, N.Oda, Y.Shimohigashi, M.Ohno, S.Hattori, M.Hattori, Y.Sakaki & H.Kihara: "Trimeresurus flavoviridis Venom Phospholipase A_2 Genes Have Evolveed via Accelerated Substitutions." Journal of Molecular Recognitioni. 8. 40-46 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Shimohigashi, T.Nose, M.Ohno, Y.Ogino & T.Costa: "Human Thrombine Receptors Are Insensitive Thrombin-Like Snake Venom." Biochemistry and Molecular Biology Int.35. 415-421 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Nakashima, I.Nobuhisa, M.Deshimaru, M.Nakai, T.Ogawa, Y.Shimohigashi, Y.Fukumaki, M.Hattori, Y.Sakaki, S.Hattori & M.Ohno: "Accelerated Evolution in the Protein-Coding Regions Is Universal in Crotalinae Snake Venom Gland Phospholipase A_2" The Proceedings of national Acae my of science USA. 92. 5605-5609 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Nakai, K.Nakashima, T.Ogawa, Y.Shimohigashi, S.Hattoti, C.C.Chang & Ohno: "Purification and Primary Structure of a Myotoxic Lysine-49-Phospholipase A_2 with Low Lipolytic Activity from Trimeresurus gramineus Venom." Toxicon. 33. 1469-1478 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Shimohigashi, A.Tani, H.Matsumoto, K.Nakashima, Y.Yamaguchi, N.Oda, Y.Takano, , H.Kamiya, J.Kishino, H.Arita & M.Ohno: "Lysine-49-Phospholipase A_2 from Trimeresurus flavoviridis Venom Are Membrane-Acting Enzymes." Journal of Biochemistry. 118. 1037-1044 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Ogawa, H,Onoue, K.Nakagawa, S.Nomura, K.Sueishi, S.Hattori, H.Kihara & M.Ohno: "Localization and Expression of Phospholipase A_2 in Trimeresurus flavoviridis (Habu Sndke) Venom Gland." Toxicon. 33. 1645-1652 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Ogawa, M.Kitajima, K.Nakashima, Y.Sakaki & M.Ohno: "Molecular Evolution of Group II Phospholipase A_2." Journal of Molecular Evolution. 41. 867-877 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] S.Nakamura, M.Nakai, K.Nakashima, T.Ogawa, Y.Shimohigashi, H.Kihara, T.Yamane, T.shida & M.Ohno: "Roles of Lysine-69 in Dimerization and Activity of Trimeresurus flavoviridis Venom Asparate 49-Phospholipase A_2." Journal of Molecular Recognition. in press (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] I.Nobuhisa, K.Nakashima, M.Deshimaru, T.Ogawa, Y.Shimohigashi, Y.Fukumaki, Y.Sakaki, S.Hattori, H.Kihara & M.Ohno: "Accelerated Evolution of Trimeresurus Okinavensis Venom Gland phospholipase A_2 Isozymes Genes." Gene. 153(in press). (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Ogawa, K.Nakashima, I.Nobuhisa, M.Deshimaru, Y.Shimohigashi, Y.Fukumaki, Y.Sakaki, S.Hattori & M.Ohno: "Accelerated Evolution of Snake Venom Phospholipase A_2 Isozymes for Acquisition of Diverse Physiological Functions." Toxicon. 34(in press). (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] B.Maillere, G.Mourier, J.Cotton, M.Herve, S.Leroy & A.Menez: "Probing immunogenicity of a T cell cpitope by L-alanine and D-amino acid scanning." Mol Immunol. 32. 1073-1080 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Dauplais, B.Gilquin, L.D.Possani, G.Gurrola-Briones, C.Roumestand & A.Menez: "Determination of the three-dimensional solution structure of noxiustoxin : analysis of structural differences with related short-chain scorpion toxins." Biochemistry. 34. 16563-16573 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] P.Cuniasse, A.Thomas, J.C.Smith, H.L.Thanh, M.Lconetti & A.Menez: "Structural basis of antibody cross-reactivity : solution conformation of an immunogenic peptide fragment containing both T and B epitopes." Biochemistry. 34. 12782-12789 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] B.Maillere, G.Mourier, M.Herve, J.Cotton, S.Leroy & A.Menez: "Immunogenicity of a disulphide-containing neurotoxin : presentation to T-cells requires a reduction step." Toxicon. 33. 475-482 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] A.L.Harvey, H.Vatanpour, E.G.Rowan, S.Pinkasfeld, C.Vita, A.Menez & M.F.Martin-Eauclaire: "Structure-activity studies on scorpion toxins that block potassium channels." Toxicon. 33. 425-436 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] R.Genet, P.H.Benetti, A.Hammadi & A.Menez: "L-tryptophan 2', 3'-oxidase from Chromobacterium violaceum.Substrate specificity and mechanistic implications." J Biol Chem. 270. 23540-23545 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] C.Vita, C.Roumestand, F.Toma & A.Menez: "Scorpion toxin as natural scaffolds for protein engineering." Proc Natl Acad Sci U S A. 92. 6404-6408 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Leonetti, J.Cotton, S.Leroy, G.Mourier & A.Menez: "Immunogenicity of T epitope-containing cyclic peptides. Increasing neutralizing antibody responses by introducing fine chemical changes." J Immunol. 155. 210-218 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Dauplais, J.M.Neumann, S.Pinkasfeld, A.Menez & C.Roumestand: "An NMR study of the interaction of cardiotoxin gamma from Naja nigricollis with perdeuterated dodecylphosphocholine micelles." Eur J Biochem. 230. 213-220 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] A.Carrier, F.Ducancel, N.B.Settiawan, L.Cattolico, B.Maillere, M.Leonetti, P.Drevet, A.Menez & J.C.Boulain: "Recombinant antibody-alkaline phosphatase conjugates for diagnosis of human IgGs : application to anti-HBsAg detection." J Immunol Methods. 181. 177-186 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] O.Tremeau, C.Lemaire, P.Drevet, S.Pinkasfeld, F.Ducancel, j.C.Boulain & A.Menez: "Genetic engineering of snake toxins.The functional site of Erabutoxin a, as delineated by site-directed mutagenesis, includes variant residues." J Biol Chem. 270. 9362-9369 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] D.Servent, A.Menez & P.Kssler: "Site-directed disulfide reduction using an affinity reagent : application on the nicotinic acetylcholine receptor." FEBS Lett.360. 261-265 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] I.Segalas, C.Roumestand, S.Zinn-Justin, B.Gilquin, R.Menez, A.Menez & F.Toma: "Solution structure of a green mamba toxin that activates muscarinic acetylcholine recptors, as studied by nuclear magnetic resonance and molecular modeling." Biochemistry. 34. 1248-1260 (1995)

    • Description
      「研究成果報告書概要(欧文)」より

URL: 

Published: 1997-03-04  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi