Improved method for the isolation, characterization and examination of neuromuscular and toxic properties of selected polypeptide fractions from the crude venom of the Taiwan cobra Naja naja atra

L. Ständker, W.G. Forssmann, L. Béress, A.L. Harvey, S. Fürst, I. Mathes, G. Escalona de Motta

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

An improved chromatographic method was developed to isolate and purify polypeptides and proteins from the crude venom of the Taiwan cobra Naja naja atra. The procedure devised is simple, easy to reproduce, and enables large scale isolation of almost all polypeptides and proteins in this cobra venom. Six pure polypeptide fractions of the venom were isolated and characterized using gel filtration on Sephadex G50 (medium), ion exchange chromatography on SP-Sephadex C25, desalting on Sephadex G25 (fine) and preparative HPLC on a RPC 18 column. The neuromuscular activity of these fractions was tested on the chick biventer cervicis nerve-muscle preparation and their toxicity (LD ) was determined after i.v. administration in mice. Their antinociceptive activity was tested in the mouse abdominal test by i.v. application. Two of these polypeptide samples had major physiological effects: one acted as a cardiotoxin causing reversible myocardial contractures with no effect on muscle twitches elicited by nerve stimulation (NS); another was a neurotoxin that blocked muscle contractions in response to NS and exogenously added acetylcholine. The cardiotoxic fraction was identified as CTX I, a well-known cardiotoxin present in this venom, and the neurotoxin was identified as neurotoxin-α with an LD50 in mice of 0.075 mg/kg.
Original languageEnglish
Pages (from-to)623-631
Number of pages9
JournalToxicon
Volume60
Issue number4
DOIs
Publication statusPublished - 15 Sep 2012

Fingerprint

Cobra Venoms
Elapidae
Poisons
Venoms
Taiwan
Neurotoxins
Cardiotoxins
Muscle
Peptides
Salt removal
Neuromuscular Junction
Lethal Dose 50
Ion Exchange Chromatography
Contracture
Muscle Contraction
Chromatography
Acetylcholine
Gel Chromatography
Toxicity
Ion exchange

Keywords

  • isolation
  • improved method
  • characterization
  • examination
  • Naja naja atra
  • neuromuscular
  • toxic properties
  • polypeptide fractions
  • crude venom
  • taiwan cobra

Cite this

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title = "Improved method for the isolation, characterization and examination of neuromuscular and toxic properties of selected polypeptide fractions from the crude venom of the Taiwan cobra Naja naja atra",
abstract = "An improved chromatographic method was developed to isolate and purify polypeptides and proteins from the crude venom of the Taiwan cobra Naja naja atra. The procedure devised is simple, easy to reproduce, and enables large scale isolation of almost all polypeptides and proteins in this cobra venom. Six pure polypeptide fractions of the venom were isolated and characterized using gel filtration on Sephadex G50 (medium), ion exchange chromatography on SP-Sephadex C25, desalting on Sephadex G25 (fine) and preparative HPLC on a RPC 18 column. The neuromuscular activity of these fractions was tested on the chick biventer cervicis nerve-muscle preparation and their toxicity (LD ) was determined after i.v. administration in mice. Their antinociceptive activity was tested in the mouse abdominal test by i.v. application. Two of these polypeptide samples had major physiological effects: one acted as a cardiotoxin causing reversible myocardial contractures with no effect on muscle twitches elicited by nerve stimulation (NS); another was a neurotoxin that blocked muscle contractions in response to NS and exogenously added acetylcholine. The cardiotoxic fraction was identified as CTX I, a well-known cardiotoxin present in this venom, and the neurotoxin was identified as neurotoxin-α with an LD50 in mice of 0.075 mg/kg.",
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Improved method for the isolation, characterization and examination of neuromuscular and toxic properties of selected polypeptide fractions from the crude venom of the Taiwan cobra Naja naja atra. / Ständker, L.; Forssmann, W.G.; Béress, L.; Harvey, A.L.; Fürst, S.; Mathes, I.; Escalona de Motta, G.

In: Toxicon, Vol. 60, No. 4, 15.09.2012, p. 623-631.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Improved method for the isolation, characterization and examination of neuromuscular and toxic properties of selected polypeptide fractions from the crude venom of the Taiwan cobra Naja naja atra

AU - Ständker, L.

AU - Forssmann, W.G.

AU - Béress, L.

AU - Harvey, A.L.

AU - Fürst, S.

AU - Mathes, I.

AU - Escalona de Motta, G.

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N2 - An improved chromatographic method was developed to isolate and purify polypeptides and proteins from the crude venom of the Taiwan cobra Naja naja atra. The procedure devised is simple, easy to reproduce, and enables large scale isolation of almost all polypeptides and proteins in this cobra venom. Six pure polypeptide fractions of the venom were isolated and characterized using gel filtration on Sephadex G50 (medium), ion exchange chromatography on SP-Sephadex C25, desalting on Sephadex G25 (fine) and preparative HPLC on a RPC 18 column. The neuromuscular activity of these fractions was tested on the chick biventer cervicis nerve-muscle preparation and their toxicity (LD ) was determined after i.v. administration in mice. Their antinociceptive activity was tested in the mouse abdominal test by i.v. application. Two of these polypeptide samples had major physiological effects: one acted as a cardiotoxin causing reversible myocardial contractures with no effect on muscle twitches elicited by nerve stimulation (NS); another was a neurotoxin that blocked muscle contractions in response to NS and exogenously added acetylcholine. The cardiotoxic fraction was identified as CTX I, a well-known cardiotoxin present in this venom, and the neurotoxin was identified as neurotoxin-α with an LD50 in mice of 0.075 mg/kg.

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KW - characterization

KW - examination

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KW - neuromuscular

KW - toxic properties

KW - polypeptide fractions

KW - crude venom

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M3 - Article

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JO - Toxicon

JF - Toxicon

SN - 0041-0101

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