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M J Dufton

Publications and source records attributed to M J Dufton.

33 records · Page 2Linked to original sources
PubMed1986

Conformational variations amongst scorpion toxins.

Circular dichroism spectra were obtained for ten scorpion neurotoxins (representing five species of scorpion) in order to provide an understanding of their relative conformations in solution. Despite a high degree of amino acid sequence homology, the toxins clearly differ from each other in terms of CD-detectable structure. When superimposed, the CD spectra suggest that the toxins form a series of related conformational variants. Since the resemblances amongst the individual CD spectra can be correlated to degrees of sequence resemblance and pharmacological specificity, conformational balance could be an important factor in both toxin evolution and target recognition.

Animals↗
PubMed1985

Proteinase inhibitors and dendrotoxins. Sequence classification, structural prediction and structure/activity.

The amino acid sequences of four presynaptically active toxins from mamba snake venom (termed 'dendrotoxins') were compared systematically with homologous sequences of members of the proteinase inhibitor family (Kunitz). A comparison based on the complete sequences revealed that relatively few amino acid changes are necessary to abolish antiprotease activity and convert a proteinase inhibitor into a dendrotoxin. When comparison centred only on the sequence segments known to comprise the antiprotease site of bovine pancreatic trypsin inhibitor, the dendrotoxins were clearly classified apart from all the known inhibitors. Since the mode of action of the bovine pancreatic trypsin/kallikrein inhibitor involves beta sheet formation with the enzyme, predictions were obtained for this secondary structure in the region of the 'antiprotease site' throughout the homologues. Again, the dendrotoxins were clearly distinguished from the inhibitors. Structure/activity analyses, based on the crystal structures of inhibitor/enzyme complexes, suggest that unlike proteinase inhibitors, dendrotoxins might specifically co-ordinate the active-site 'catalytic' histidine residues of serine proteases. Although the significance of this remains to be studied, the presynaptic target is expected to involve an as yet uncharacterised member of the serine protease family.

Amino Acid Sequence↗
PubMed1985

Genetic code redundancy and the evolutionary stability of protein secondary structure.

The genetic code has an inherent bias towards some amino acids because of the variable number of synonymous codons per amino acid. The extent to which these biases are expressed in protein secondary structure is described through the analysis of the overall amino acid compositions of the alpha-helix, beta-sheet, beta-turn and random coil segments elucidated by X-ray crystallography. Given the concept of neutral mutation in proteins, the allocation of synonyms in the genetic code appears to protect secondary structures from amino acid changes and discourages the appearance of chemically complex residues. The level of protection is similar for each structural form, despite their clear preferences for certain amino acids. The organization of the code is therefore relevant to the preservation of conformation seen in the evolution of many protein families.

Amino Acids↗
PubMed1985

Protein rotational diffusion measurements on the interaction of bee venom melittin with bacteriorhodopsin in lipid vesicles.

The rotational diffusion of bacteriorhodopsin reconstituted into dimyristoylphosphatidylcholine vesicles was measured by the technique of flash-induced transient dichroism. In the presence of melittin, a cell lysing peptide from honey bee (Apis mellifera) venom, dose-dependent loss of rotational mobility was observed. Chemically modified melittin derivatives, in which free amine groups were either acetylated or succinylated, were impaired in their ability to induce immobilisation of bacteriorhodopsin. Bacteriorhodopsin reconstitutions of differing lipid/protein ratio were tested and it was found that the bacteriorhodopsin immobilisation phenomena depended on the melittin/protein ratio, not the melittin/lipid ratio. This suggests that melittin produces its effect via direct interaction with bacteriorhodopsin. A mechanism is proposed in which the aggregation of bacteriorhodopsin is induced by electrostatic attraction between its anionic surface moieties and the highly cationic C-terminal segment of melittin.

Acetylation↗
PubMed1984

The capacity of basic peptides to trigger exocytosis from mast cells correlates with their capacity to immobilize band 3 proteins in erythrocyte membranes.

The effect of mast-cell-triggering peptides on the rotational properties of band 3, a protein component of the human erythrocyte membrane, was measured by observing flash-induced transient dichroism of the triplet probe eosin maleimide. In the presence of melittin, polylysine and five synthetic peptides, varying degrees of retardation in the rotational motion of band 3 were produced. When placed in order of band 3 immobilizing activity, the peptides formed a series identical with their order of efficacy in releasing 5-hydroxytryptamine from rat peritoneal mast cells. The correspondence in the abilities to immobilize band 3 in the erythrocyte and trigger mast cells is significant because structure-activity analyses of the peptides show both processes to have the same cationic, hydrophobic and stereochemical requirements. Probably, the immobilization of band 3 proteins reflects an ability of the basic peptides to aggregate anionic surface moieties, and therefore a similar mechanism is implied in mast-cell triggering.

Animals↗
PubMed1984

The influence of melittin on the rotation of band 3 protein in the human erythrocyte membrane.

The rotational mobility of band 3, a protein constituent of the human erythrocyte membrane, was measured by observing the flash-induced transient dichroism of the triplet probe eosin maleimide. In the presence of melittin, a pharmacologically active polypeptide from honey bee (Apis mellifera) venom, a dose-dependent loss of rotational mobility was detected. With acetylated melittin, the ability to immobilize is reduced. Succinylated melittin, however, is devoid of immobilizing activity. The possible relevance of these findings to the normal mode of action of melittin was examined by comparing the relative abilities of the native, acetylated and succinylated melittins to lyse erythrocytes and synergize with phospholipase A2, another constituent of bee venom. For both these properties, the order of effectiveness is native melittin greater than acetyl melittin greater than succinyl melittin = 0, the same as their order of effectiveness in immobilizing band 3. A mechanism is proposed in which melittin is anchored in the membrane by its hydrophobic N-terminus, while its cationic C-terminal moiety binds to negatively charged residues on membrane proteins. This leads either directly or indirectly to protein aggregation and hence loss of mobility. From a detailed comparison of the different effects of the melittin derivatives, it is concluded that melittin may function in vivo by aggregating membrane proteins in order to allow phospholipase A2 to gain access to the membrane bilayer and commence catalysis.

Anion Exchange Protein 1, Erythrocyte↗
PubMed1984

Classification of scorpion toxins according to amino acid composition and sequence.

Scorpion venom toxins were systematically classified according to amino acid composition, insertion/deletion events and sequence. The significance of each comparison method and its outcome is discussed in relation to known immunological and structural properties. A general classification of the toxins is proposed that accounts for both the immunological groupings and the differences in mode of action.

Amino Acid Sequence↗
PubMed1984

Classification of elapid snake neurotoxins and cytotoxins according to chain length: evolutionary implications.

The amino acid sequences of the 139 homologous "short" neurotoxins, "long" neurotoxins and cytotoxins so far characterised from elapid snake venoms were compared on the basis of the amino acid deletion/insertion events that have occurred during evolution. Systematic grouping of the toxins according to similarity suggests that the short neurotoxins resemble the cytotoxins more closely than they do the long neurotoxins. The significance of this finding is discussed in relation to the methodology, the conformations of the toxins (as represented by circular dichroism spectra) and the outcome of the study that would have been obtained. had more traditional methods been used. It appears probable that the cytotoxins evolved relatively recently from neurotoxic ancestors.

Amino Acid Sequence↗
PubMed1983

Conformational properties of phospholipases A2. Secondary-structure prediction, circular dichroism and relative interface hydrophobicity.

The sequences of 32 phospholipases A2 (EC 3.1.1.4) were analysed by secondary-structure prediction and the results were compared with the available crystallographic data. Good agreement is evident between prediction and experiment, especially for helical structure. Circular dichroic spectra were also determined for six enzymes from Elapid snake venom and these, in association with previously published spectra, confirm the main implication of the predictions, namely that all the homologues have qualitatively similar tertiary structures. Consideration was then given to possible structure/activity relationships in the light of the above findings. The relative hydrophobicity/hydrophilicity of the area of the enzyme thought to interact with lipid/water interfaces was predicted and certain correlations were noted with relative penetrating power, species of origin and the presence of beta-neurotoxic properties.

Amino Acid Sequence↗
PubMed1983

Classification of phospholipases A2 according to sequence. Evolutionary and pharmacological implications.

The sequences of 32 phospholipases A2 (EC 3.1.1.4) were systematically compared on the basis of polypeptide chain length and similarity at selected amino acid positions around the active site. Two difference matrices were constructed and the various groupings present in the data were expressed in dendrogram form. The two methods of comparison yielded different results, and this is seen as a consequence of separate aspects of phospholipase evolution being highlighted in each case. It appears that, although Elapid snake venom phospholipases are very similar in terms of overall conformation, the area around their active sites distinguishes them into two major groups, namely the Asian Elapids and the marine/Australasian Elapids. Further, the Asian Elapids seem to have active-site vicinities which are closer to those in the mammalian pancreatic phospholipases. The relevance of the classifications to structure/activity relationships (especially beta-neurotoxicity) and phospholipase evolution is discussed.

Amino Acid Sequence↗
PubMed1983

The significance of redundancy in the genetic code.

The genetic code has an inherent bias towards some amino acids because of the variable number of synonymous codons per amino acid. In proteins generally, this bias is expressed in the relative proportions of the twenty amino acids. It is suggested that even though neutral mutation may be responsible for the expression of this bias, the latter could be providing a positive advantage by directing mutation to introduce chemically simpler and more immutable amino acids where selective criteria have become relaxed.

Amino Acid Sequence↗
PubMed1983

Conformational properties of the neurotoxins and cytotoxins isolated from Elapid snake venoms.

The review will critically assess the information available on the conformation of homologous neurotoxins and cytotoxins isolated from Elapid snakes. Particular attention will be given to the dynamics of the molecules in solution because there is the possibility that defined intramolecular rearrangements are involved at the sites of action. Such properties will be then reconciled with the known X-ray crystallographic and sequence data in order to derive likely structure-activity relationships.

Amino Acid Sequence↗
PubMed1980

Circular dichroism of elapidae protein toxins.

The effect of temperature variation on the circular dichroic spectra of four Elapid snake toxins (one short neurotoxin, two long neurotoxins and one cytotoxin) was determined and the results compared. The three neurotoxins have in common a delicately balanced conformation which can be perturbed independently of the predominant secondary structuring. Despite the many differences in sequence between long and short neurotoxins, the nature of this balance seems to be the same in both types and it is considered to involve an equilibrium between two defined conformers. Such properties could not be discerned in the cytotoxin studied, but related work suggests that they could nevertheless be present in some cases. In terms of global structure, long neurotoxins may have a greater proportion of 'random chain' inherent than short neurotoxins, the latter having extensive beta-sheet. The important feature of all toxins is deemed to be a conformational freedom defined by the secondary structuring and characteristic disulphide bridge formation. Possible relevance to toxicity of such a property is discussed.

Amino Acid Sequence↗