Cobra cardiotoxins. Purification, effects on skeletal muscle and structure/activity relationships

S J Hodges, A S Agbaji, A L Harvey, R C Hider

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

A new preparative method for isolating homogeneous cardiotoxins from cobra venoms is described. The technique, based on reverse-phase high-pressure liquid chromatography, was used to isolate eight cardiotoxins of known sequence from four different venoms. In each case the method was found to be particularly efficient at removing trace quantities of contaminating phospholipase. Cardiotoxins isolated in this manner were found to retain their full biological activity. Without exception the purified cardiotoxins lacked powerful haemolytic activity at concentrations up to 0.01 mM (about 100 micrograms ml-1), although some lysis of human erythrocytes was induced at higher concentrations. The cardiotoxins displayed a wide range of depolarizing activity on cultured skeletal muscle, the lowest activity being associated with the highest LD50 value. Correlating variations in amino acid sequence and variations in depolarization potency revealed the importance of residues in the second and third loops, especially lysine-46, serine-48 and lysine-52, together with a number of hydrophobic residues. Further modifications of pharmacological activity were associated with the presence of additional basic residues in the first and second loops and to minor differences in secondary structure.
Original languageEnglish
Pages (from-to)373-383
Number of pages11
JournalEuropean Journal of Biochemistry
Volume165
Issue number2
Publication statusPublished - 1 Jun 1987

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Cobra Cardiotoxin Proteins
Cardiotoxins
Structure-Activity Relationship
Purification
Muscle
Skeletal Muscle
Lysine
High pressure liquid chromatography
Cobra Venoms
Phospholipases
Venoms
Depolarization
Bioactivity
Serine
Lethal Dose 50
Reverse-Phase Chromatography
Amino Acid Sequence
Amino Acids
Erythrocytes
High Pressure Liquid Chromatography

Keywords

  • amino acid sequence
  • amino acids
  • animals
  • chick embryo
  • chromatography
  • circular dichroism
  • cobra cardiotoxin proteins
  • cobra venoms
  • erythrocytes
  • humans
  • hydrolysis
  • isoelectric focusing
  • muscles
  • phospholipases A
  • structure-activity relationship

Cite this

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title = "Cobra cardiotoxins. Purification, effects on skeletal muscle and structure/activity relationships",
abstract = "A new preparative method for isolating homogeneous cardiotoxins from cobra venoms is described. The technique, based on reverse-phase high-pressure liquid chromatography, was used to isolate eight cardiotoxins of known sequence from four different venoms. In each case the method was found to be particularly efficient at removing trace quantities of contaminating phospholipase. Cardiotoxins isolated in this manner were found to retain their full biological activity. Without exception the purified cardiotoxins lacked powerful haemolytic activity at concentrations up to 0.01 mM (about 100 micrograms ml-1), although some lysis of human erythrocytes was induced at higher concentrations. The cardiotoxins displayed a wide range of depolarizing activity on cultured skeletal muscle, the lowest activity being associated with the highest LD50 value. Correlating variations in amino acid sequence and variations in depolarization potency revealed the importance of residues in the second and third loops, especially lysine-46, serine-48 and lysine-52, together with a number of hydrophobic residues. Further modifications of pharmacological activity were associated with the presence of additional basic residues in the first and second loops and to minor differences in secondary structure.",
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year = "1987",
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Cobra cardiotoxins. Purification, effects on skeletal muscle and structure/activity relationships. / Hodges, S J; Agbaji, A S; Harvey, A L; Hider, R C.

In: European Journal of Biochemistry , Vol. 165, No. 2, 01.06.1987, p. 373-383.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Cobra cardiotoxins. Purification, effects on skeletal muscle and structure/activity relationships

AU - Hodges, S J

AU - Agbaji, A S

AU - Harvey, A L

AU - Hider, R C

PY - 1987/6/1

Y1 - 1987/6/1

N2 - A new preparative method for isolating homogeneous cardiotoxins from cobra venoms is described. The technique, based on reverse-phase high-pressure liquid chromatography, was used to isolate eight cardiotoxins of known sequence from four different venoms. In each case the method was found to be particularly efficient at removing trace quantities of contaminating phospholipase. Cardiotoxins isolated in this manner were found to retain their full biological activity. Without exception the purified cardiotoxins lacked powerful haemolytic activity at concentrations up to 0.01 mM (about 100 micrograms ml-1), although some lysis of human erythrocytes was induced at higher concentrations. The cardiotoxins displayed a wide range of depolarizing activity on cultured skeletal muscle, the lowest activity being associated with the highest LD50 value. Correlating variations in amino acid sequence and variations in depolarization potency revealed the importance of residues in the second and third loops, especially lysine-46, serine-48 and lysine-52, together with a number of hydrophobic residues. Further modifications of pharmacological activity were associated with the presence of additional basic residues in the first and second loops and to minor differences in secondary structure.

AB - A new preparative method for isolating homogeneous cardiotoxins from cobra venoms is described. The technique, based on reverse-phase high-pressure liquid chromatography, was used to isolate eight cardiotoxins of known sequence from four different venoms. In each case the method was found to be particularly efficient at removing trace quantities of contaminating phospholipase. Cardiotoxins isolated in this manner were found to retain their full biological activity. Without exception the purified cardiotoxins lacked powerful haemolytic activity at concentrations up to 0.01 mM (about 100 micrograms ml-1), although some lysis of human erythrocytes was induced at higher concentrations. The cardiotoxins displayed a wide range of depolarizing activity on cultured skeletal muscle, the lowest activity being associated with the highest LD50 value. Correlating variations in amino acid sequence and variations in depolarization potency revealed the importance of residues in the second and third loops, especially lysine-46, serine-48 and lysine-52, together with a number of hydrophobic residues. Further modifications of pharmacological activity were associated with the presence of additional basic residues in the first and second loops and to minor differences in secondary structure.

KW - amino acid sequence

KW - amino acids

KW - animals

KW - chick embryo

KW - chromatography

KW - circular dichroism

KW - cobra cardiotoxin proteins

KW - cobra venoms

KW - erythrocytes

KW - humans

KW - hydrolysis

KW - isoelectric focusing

KW - muscles

KW - phospholipases A

KW - structure-activity relationship

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