ABSTRACT
Erabutoxins a and b are neurotoxins isolated from venom of a sea snake Laticauda semifasciata (erabu-umihebi). Amino acid sequences of the toxins indicated that the toxins are members of a superfamily consisting of short and long neurotoxins and cytotoxins found in sea snakes and terrestrial snakes. The short neurotoxins to which erabutoxins belong act by blocking the nicotinic acetylcholine receptor on the post synaptic membrane in a manner similar to that of curare. X-ray crystallography and NMR analyses showed that the toxins have a three-finger structure, in which three fingers made of three loops emerging from a dense core make a gently concave surface of the protein. The sequence comparison and the location of essential residues on the protein suggested the mechanism of binding of the toxin to the acetylcholine receptor. Classification of snakes by means of sequence comparison and that based on different morphological features were inconsistent, which led the authors to propose a hypothesis "Evolution without divergence."
Subject(s)
Elapid Venoms , Elapidae , Amino Acid Sequence , Animals , Elapid Venoms/chemistry , Elapid Venoms/isolation & purification , Elapid Venoms/metabolism , Elapid Venoms/toxicity , Elapidae/genetics , Erabutoxins/chemistry , Erabutoxins/isolation & purification , Erabutoxins/metabolism , Erabutoxins/toxicity , Evolution, Molecular , Humans , Molecular Sequence Data , Neurotoxins/chemistry , Neurotoxins/isolation & purification , Neurotoxins/metabolism , Neurotoxins/toxicity , Receptors, Cholinergic/metabolismABSTRACT
A study has been made, following high-resolution refinement at 0.14 nm, of the structure of erabutoxin b, prototype postsynaptic neurotoxin from snake venom. The detailed patterns of intramolecular van der Waal's interactions have been determined. From information, hitherto unavailable, about atomic temperature parameters, the relative mobilities in different regions of the molecule have been estimated. A detailed model of structure/function relationships in these neurotoxins, which bind to the acetylcholine receptor, has thus been established: the probable dynamic mode of toxin-receptor binding is described. The model identifies, and the binding mode depends on a unique structural feature of these protein toxins: the hydrophobic 'Trp' cleft. Charge-charge interactions are implicated in initial toxin orientation on the receptor surface. Possible reactive-site extension in short-chain toxins is described. Modifications in binding mode of long-chain toxins are considered. The relative mobilities of antigenic site residues are discussed.
Subject(s)
Elapid Venoms/isolation & purification , Erabutoxins/isolation & purification , Binding Sites , Epitopes , Erabutoxins/immunology , Erabutoxins/metabolism , Models, Molecular , Protein Conformation , Receptors, Cholinergic/metabolism , Structure-Activity RelationshipABSTRACT
Erabutoxins a and b, the major neurotoxins in the venom of the sea snake Laticauda semifasciata, were detected in the venom of Laticauda schistorhynchus. The identity of the toxins was confirmed on the basis of elution position on CM-cellulose column chromatography, disc electrophoretic mobility, amino acid analysis and toxicity measurement.
Subject(s)
Elapid Venoms , Elapid Venoms/analysis , Erabutoxins , Neurotoxins , Amino Acids/analysis , Animals , Chemical Phenomena , Chemistry , Chromatography, Ion Exchange , Elapid Venoms/isolation & purification , Erabutoxins/isolation & purification , Neurotoxins/isolation & purification , Species SpecificityABSTRACT
1. Erabutoxins, a, b and c, neurotoxic proteins of a sea snake Lacticauda semifasciata, were guanidinated with O-methylisourea. The amino groups of all the lysine residues and those at the N-termini of the toxins were modified. The lethal activity of the toxins decreased to 50% (erabutoxins a and b) or 17% (erabutoxin c) of the original value on the modification. The c.d. (circular dichroism) maximum at 227 nm of the modified toxins became lower, whereas the whole profile of the c.d. curve remained unchanged. 2. The amino groups of erabutoxin b were acetylated with acetic anhydride. All the five monoacetyl derivatives were isolated from the reaction products by CM-cellulose and Bio-Rex 70 column chromatography. [1-Nalpha-acetylarginine]-, [15-N6-acetyl-lysine]- and [51-N6-acetyl-lysine]-erabutoxin b retained the toxicity of the native toxin, whereas [27-N6-acetyl-lysine] and [47-N6-acetyl-lysine]-erabutoxin b were 17 and 8% active respectively. The overall profile of c.d. spectrum of erabutoxin b remained unchanged on the monoacetylation.
