On the purification of notexin: Isolation of a single amino acid variant from the venom of Notechis scutatus scutatus
Venom of the Australian tiger snake, Notechis scutatus scutatus was fractionated by conventional ion-exchange chromatography. The fraction containing notexin, a well-known single-chain toxic phospholipase A 2, was further purified by reverse-phase high-performance liquid chromatography. Two main com...
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| Published in: | FEBS letters Vol. 261; no. 2; pp. 226 - 230 |
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| Main Authors: | , , , , , |
| Format: | Journal Article |
| Language: | English |
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Elsevier B.V
26-02-1990
Elsevier Wiley |
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| Abstract | Venom of the Australian tiger snake,
Notechis scutatus scutatus was fractionated by conventional ion-exchange chromatography. The fraction containing notexin, a well-known single-chain toxic phospholipase A
2, was further purified by reverse-phase high-performance liquid chromatography. Two main components were isolated and the major one corresponded to notexin. The other component, designated as notechis N
s, was an isofonn of notexin. Notechis N
s and notexin possessed similar in vitro esterase activity, in vitro neuromuscular activity and in vivo lethality. Amino acid composition and sequence of the
Staphylococcus aureus V8-protease peptides demonstrated that primary structures of notechis N
s and notexin differed from each other by a single substitution amongst 119 amino acids: Lys → Arg at position 16. |
|---|---|
| AbstractList | Venom of the Australian tiger snake, Notechis scutatus scutatus was fractionated by conventional ion-exchange chromatography. The fraction containing notexin, a well-known single-chain toxic phospholipase A2, was further purified by reverse-phase high-performance liquid chromatography. Two main components were isolated and the major one corresponded to notexin. The other component, designated as notechis Ns, was an isoform of notexin. Notechis Ns and notexin possessed similar in vitro esterase activity, in vitro neuromuscular activity and in vivo lethality. Amino acid composition and sequence of the Staphylococcus aureus V8-protease peptides demonstrated that primary structures of notechis Ns and notexin differed from each other by a single substitution amongst 119 amino acids: Lys---Arg at position 16. Venom of the Australian tiger snake, Notechis scutatus scutatus was fractionated by conventional ion-exchange chromatography. The fraction containing notexin, well-known single-chain toxic phospholipase A sub(2), was further purified by reverse-phase high-performance liquid chromatography. Two main components were isolated and the major one corresponded to notexin. The other component, designated as notechis N sub(s), was an isoform of notexin. Notechis N sub(s) and notexin possessed similar in vitro esterase activity, in vitro neuromuscular activity and in vivo lethality. Amino acid composition and sequence of the Staphylococcus aureus) V8-protease peptide demonstrated that primary structures of notechis N sub(s) and notexin differed from each other by a single substitution amongst 119 amino acids. Venom of the Australian tiger snake, Notechis scutatus scutatus was fractionated by conventional ion-exchange chromatography. The fraction containing notexin, a well-known single-chain toxic phospholipase A 2, was further purified by reverse-phase high-performance liquid chromatography. Two main components were isolated and the major one corresponded to notexin. The other component, designated as notechis N s, was an isofonn of notexin. Notechis N s and notexin possessed similar in vitro esterase activity, in vitro neuromuscular activity and in vivo lethality. Amino acid composition and sequence of the Staphylococcus aureus V8-protease peptides demonstrated that primary structures of notechis N s and notexin differed from each other by a single substitution amongst 119 amino acids: Lys → Arg at position 16. Venom of the Australian tiger snake, Notechis scutatus scutatus was fractionated by conventional ion‐exchange chromatography. The fraction containing notexin, a well‐known single‐chain toxic phospholipase A 2 , was further purified by reverse‐phase high‐performance liquid chromatography. Two main components were isolated and the major one corresponded to notexin. The other component, designated as notechis N s , was an isofonn of notexin. Notechis N s and notexin possessed similar in vitro esterase activity, in vitro neuromuscular activity and in vivo lethality. Amino acid composition and sequence of the Staphylococcus aureus V8‐protease peptides demonstrated that primary structures of notechis N s and notexin differed from each other by a single substitution amongst 119 amino acids: Lys → Arg at position 16. |
| Author | Harvey, Alan L. Rowan, Edward G. Mollier, Pascale Ménez, André Chwetzoff, Serge Bouet, Françoise |
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| Keywords | Reverse-phase high-performance liquid chromatography Phospholipase A 2 LD 50, median lethal dose Notexin Isoform EPPS, end-plate potentials MEPPS, miniature end-plate potentials RP-HPLC, reverse-phase high-performance liquid chromatography Vertebrata Purification Enzyme Genetic variant Venom HPLC chromatography Isolation Reptilia Phospholipase A Ion exchange chromatography Ophidia |
| Language | English |
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| References | Crestfield, Moore, Stein (BIB11) 1963; 238 Harris, Johnson (BIB2) 1978; 5 Howard, Gundersen (BIB7) 1980; 20 Chang (BIB9) 1985; 9 Mollier, Chwetzoff, Bouet, Harvey, Ménez (BIB10) 1989; 185 Harris, Karlsson, Thesleff (BIB1) 1973; 47 Halpert, Eaker (BIB5) 1975; 250 Gueriani-Gruszka, Almag, Biltonen, Lichtenberg (BIB14) 1988; 263 Mollier (BIB15) 1987 Lind, Eaker (BIB4) 1980; 111 Dixon, Mood (BIB12) 1948; 43 Harris (BIB8) 1984; 21 Karlsson, Eaker, Ryden (BIB6) 1972; 10 Rowan, Harvey (BIB13) 1988; 94 Halpert, Eaker (BIB3) 1976; 251 Harris J.B. (e_1_2_1_9_1) 1984; 21 Chang C.C. (e_1_2_1_10_1) 1985; 9 e_1_2_1_7_1 e_1_2_1_8_1 Mollier P. (e_1_2_1_16_1) 1987 e_1_2_1_6_1 e_1_2_1_3_1 e_1_2_1_12_1 e_1_2_1_4_1 e_1_2_1_13_1 e_1_2_1_2_1 e_1_2_1_11_1 e_1_2_1_14_1 e_1_2_1_15_1 Lind P. (e_1_2_1_5_1) 1980; 111 |
| References_xml | – volume: 111 start-page: 403 year: 1980 end-page: 409 ident: BIB4 publication-title: Bur. J. Biochem. contributor: fullname: Eaker – volume: 94 start-page: 839 year: 1988 end-page: 847 ident: BIB13 publication-title: Br. J. Pharmacol. contributor: fullname: Harvey – volume: 5 start-page: 587 year: 1978 end-page: 600 ident: BIB2 publication-title: Clin. Exp. Pharmacol. Physiol. contributor: fullname: Johnson – volume: 238 start-page: 622 year: 1963 end-page: 627 ident: BIB11 publication-title: J. Biol. Chem. contributor: fullname: Stein – volume: 185 start-page: 263 year: 1989 end-page: 270 ident: BIB10 publication-title: Eur. J. Biochem. contributor: fullname: Ménez – volume: 251 start-page: 7343 year: 1976 end-page: 7445 ident: BIB3 publication-title: J. Biol. Chem. contributor: fullname: Eaker – volume: 10 start-page: 405 year: 1972 end-page: 413 ident: BIB6 publication-title: Toxicon contributor: fullname: Ryden – volume: 21 start-page: 64 year: 1984 end-page: 110 ident: BIB8 publication-title: Prog. Med. Chem. contributor: fullname: Harris – volume: 263 start-page: 11808 year: 1988 end-page: 11813 ident: BIB14 publication-title: J. Biol. Chem. contributor: fullname: Lichtenberg – volume: 250 start-page: 6990 year: 1975 end-page: 6997 ident: BIB5 publication-title: J. Biol. Chem. contributor: fullname: Eaker – volume: 20 start-page: 307 year: 1980 end-page: 336 ident: BIB7 publication-title: Annu. Rev. Pharmacol. Toxicol. contributor: fullname: Gundersen – volume: 43 start-page: 109 year: 1948 end-page: 126 ident: BIB12 publication-title: J. Am. Stat. Assoc. contributor: fullname: Mood – year: 1987 ident: BIB15 publication-title: Thèse de l'Université Paris XI contributor: fullname: Mollier – volume: 9 start-page: 126 year: 1985 end-page: 142 ident: BIB9 publication-title: Proc. Natl. Acad. Counc. Repub. China part B Life. Sci. contributor: fullname: Chang – volume: 47 start-page: 141 year: 1973 end-page: 146 ident: BIB1 publication-title: Br. J. Pharmacol. contributor: fullname: Thesleff – ident: e_1_2_1_11_1 doi: 10.1111/j.1432-1033.1989.tb15111.x – year: 1987 ident: e_1_2_1_16_1 publication-title: Thèse de l'Université Paris XI contributor: fullname: Mollier P. – ident: e_1_2_1_12_1 doi: 10.1016/S0021-9258(18)81308-4 – ident: e_1_2_1_4_1 doi: 10.1016/S0021-9258(17)32855-7 – ident: e_1_2_1_13_1 doi: 10.1080/01621459.1948.10483254 – volume: 9 start-page: 126 year: 1985 ident: e_1_2_1_10_1 publication-title: Proc. Natl. Acad. Counc. Repub. China part B Life. Sci. contributor: fullname: Chang C.C. – ident: e_1_2_1_3_1 doi: 10.1111/j.1440-1681.1978.tb00714.x – ident: e_1_2_1_6_1 doi: 10.1016/S0021-9258(19)41030-2 – ident: e_1_2_1_14_1 doi: 10.1111/j.1476-5381.1988.tb11595.x – ident: e_1_2_1_15_1 doi: 10.1016/S0021-9258(18)37857-8 – volume: 111 start-page: 403 year: 1980 ident: e_1_2_1_5_1 publication-title: Bur. J. Biochem. contributor: fullname: Lind P. – ident: e_1_2_1_2_1 doi: 10.1111/j.1476-5381.1973.tb08168.x – ident: e_1_2_1_7_1 doi: 10.1016/0041-0101(72)90066-9 – ident: e_1_2_1_8_1 doi: 10.1146/annurev.pa.20.040180.001515 – volume: 21 start-page: 64 year: 1984 ident: e_1_2_1_9_1 publication-title: Prog. Med. Chem. contributor: fullname: Harris J.B. |
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| Snippet | Venom of the Australian tiger snake,
Notechis scutatus scutatus was fractionated by conventional ion-exchange chromatography. The fraction containing notexin,... Venom of the Australian tiger snake, Notechis scutatus scutatus was fractionated by conventional ion-exchange chromatography. The fraction containing notexin,... Venom of the Australian tiger snake, Notechis scutatus scutatus was fractionated by conventional ion‐exchange chromatography. The fraction containing notexin,... |
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| SubjectTerms | Amino Acid Sequence Analytical, structural and metabolic biochemistry Animals Biological and medical sciences Biological Assay Chemical Sciences Chickens Chromatography, High Pressure Liquid Chromatography, Ion Exchange Elapid Venoms - isolation & purification Elapid Venoms - pharmacology Elapid Venoms - toxicity Enzymes and enzyme inhibitors Esterases - metabolism Female Fundamental and applied biological sciences. Psychology Hydrolases Hydrolysis Isoform Lethal Dose 50 Mice Mice, Inbred BALB C Molecular Sequence Data Neuromuscular Junction - drug effects Neuromuscular Junction - physiology Notechis scutatus scutatus Notexin Organic chemistry Peptide Fragments Phospholipase A 2 Rana pipiens Reverse-phase high-performance liquid chromatography Synaptic Transmission - drug effects |
| Title | On the purification of notexin: Isolation of a single amino acid variant from the venom of Notechis scutatus scutatus |
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