On the purification of notexin: Isolation of a single amino acid variant from the venom of Notechis scutatus scutatus

• Published in: FEBS letters Vol. 261; no. 2; pp. 226 - 230
• Main Authors: Chwetzoff, Serge, Mollier, Pascale, Bouet, Françoise, Rowan, Edward G., Harvey, Alan L., Ménez, André
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 26-02-1990; Elsevier; Wiley
• Subjects: 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; 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
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(90)80559-2

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.